1 00:00:05,240 --> 00:00:07,000 WELCOME TO THE COVID-19 2 00:00:07,000 --> 00:00:09,560 SCIENTIFIC INTEREST GROUP 3 00:00:09,560 --> 00:00:10,680 LECTURE SERIES. 4 00:00:10,680 --> 00:00:13,960 THIS IS THE NO GROW LECTURE FOR 5 00:00:13,960 --> 00:00:17,440 THE NEW YEAR, 2022. 6 00:00:17,440 --> 00:00:20,720 WE ARE DELIGHTED TO HAVE DR. 7 00:00:20,720 --> 00:00:22,920 MELANIE OTT KICKING IT OFF FOR 8 00:00:22,920 --> 00:00:23,400 US. 9 00:00:23,400 --> 00:00:25,960 DR. OTT IS THE DIRECTOR OF THE 10 00:00:25,960 --> 00:00:29,080 GLADSTONE INSTITUTE OF VIROLOGY 11 00:00:29,080 --> 00:00:31,600 AND PROFESSOR OF MEDICINE AT 12 00:00:31,600 --> 00:00:32,520 UCSF. 13 00:00:32,520 --> 00:00:35,640 SHE'S A WORLDWIDE LEADER IN 14 00:00:35,640 --> 00:00:37,160 INVESTIGATIONS INTO THE HOST 15 00:00:37,160 --> 00:00:40,440 VIRUS INTERFACE. 16 00:00:40,440 --> 00:00:42,160 STUDYING BOTH VIRAL LIFE CYCLE 17 00:00:42,160 --> 00:00:43,640 AND USING VIRUSES AS TOOLS TO 18 00:00:43,640 --> 00:00:45,880 MAKE DISCOVERIES ABOUT THE INNER 19 00:00:45,880 --> 00:00:46,680 WORKING CELL. 20 00:00:46,680 --> 00:00:48,880 SHE'S MADE IMPORTANT DISCOVERIES 21 00:00:48,880 --> 00:00:51,080 ABOUT HOW VIRUSES INCLUDING 22 00:00:51,080 --> 00:00:55,680 HEPATITIS C ZIKA, HIV AND TODAY 23 00:00:55,680 --> 00:00:59,080 YOU WILL HEAR COVID, HIJACK 24 00:00:59,080 --> 00:00:59,680 HUMAN CELLS. 25 00:00:59,680 --> 00:01:02,880 HER STUDIES ON HIV IN PARTICULAR 26 00:01:02,880 --> 00:01:06,040 ABOUT LATENCY PHASE LED TO 27 00:01:06,040 --> 00:01:09,760 SIMILAR KNOLL DISCOVERIES HOW 28 00:01:09,760 --> 00:01:11,440 PROTEINS MANIPULATE THE 29 00:01:11,440 --> 00:01:13,880 EPIGENETIC CODE CHANGE CHROMATIN 30 00:01:13,880 --> 00:01:15,520 ORGANIZATION AND CONTRIBUTE TO 31 00:01:15,520 --> 00:01:18,160 PREMATURE IMMUNE CELL AGING BY 32 00:01:18,160 --> 00:01:18,880 MANIPULATING CIRCUMSTANCE 33 00:01:18,880 --> 00:01:19,440 PROTEINS. 34 00:01:19,440 --> 00:01:21,920 HER WORK ON HCV LED TO DEEPER 35 00:01:21,920 --> 00:01:23,920 UNDERSTANDING HOW VIRUSES 36 00:01:23,920 --> 00:01:25,600 REGULAR RATE THE LIPID 37 00:01:25,600 --> 00:01:29,240 METABOLISM OF CELLS AND HER WORK 38 00:01:29,240 --> 00:01:31,880 ON ZIKA AND HCV REVEAL 39 00:01:31,880 --> 00:01:33,320 MECHANISMS WHICH VIRUSES 40 00:01:33,320 --> 00:01:36,920 MODULATE RNA METABOLISM OF CELL 41 00:01:36,920 --> 00:01:39,680 PERTURBING NON-SENSE MEDIATED 42 00:01:39,680 --> 00:01:42,560 mRNA PATHWAYS AND RNA IMPORT 43 00:01:42,560 --> 00:01:44,120 EXPORT PATHWAYS. 44 00:01:44,120 --> 00:01:46,560 SHE ALSO IS TOGETHER WITH 45 00:01:46,560 --> 00:01:49,120 JENNIFER AND DAN FLETCHER HAS 46 00:01:49,120 --> 00:01:50,480 BEEN DEVELOPING CRISPER BASED 47 00:01:50,480 --> 00:01:52,680 VIRUS TESTING PLATFORMS THAT DO 48 00:01:52,680 --> 00:01:55,360 NOT NEED TO BE CARRIED OUT IN A 49 00:01:55,360 --> 00:01:57,880 LAB USE MOBILE PHONE 50 00:01:57,880 --> 00:02:00,040 TECHNOLOGIES, AND DELIVER 51 00:02:00,040 --> 00:02:00,880 QUANTITATIVE RESULTS IN SHORT 52 00:02:00,880 --> 00:02:03,120 AMOUNTS OF TIME. 53 00:02:03,120 --> 00:02:04,840 MOST RECENTLY LIKE THE REST OF 54 00:02:04,840 --> 00:02:07,240 THE WORLD HER FOCUSES TURN TO 55 00:02:07,240 --> 00:02:11,080 SARS COV-2 AND TODAY YOU WILL BE 56 00:02:11,080 --> 00:02:12,680 HEARING ABOUT SOME RECENT WORK 57 00:02:12,680 --> 00:02:17,920 THAT JUST CAME OUT IN SCIENCE 58 00:02:17,920 --> 00:02:20,880 WHERE SHE AND DR. CONNER 59 00:02:20,880 --> 00:02:22,200 DEVELOPED COV-2 VIRUS LIKE 60 00:02:22,200 --> 00:02:24,120 PARTICLES TO LOOK AT IMPACT OF 61 00:02:24,120 --> 00:02:25,880 MUTATIONS ON N AND SPIKE 62 00:02:25,880 --> 00:02:28,040 PROTEINS ON NOT ONLY VIRUS ENTRY 63 00:02:28,040 --> 00:02:29,920 BUT INTERESTINGLY RNA RELEASE 64 00:02:29,920 --> 00:02:33,000 AND RNA PACKAGING. 65 00:02:33,000 --> 00:02:34,360 SO WE HOPE TO HEAR ABOUT SOME OF 66 00:02:34,360 --> 00:02:36,040 THAT TODAY. 67 00:02:36,040 --> 00:02:36,880 WITHOUT FURTHER ADIEU, I WOULD 68 00:02:36,880 --> 00:02:39,840 LIKE TO WELCOME DR. OTT TO NIH, 69 00:02:39,840 --> 00:02:40,680 WE LOOK FORWARD TO HEARING HER 70 00:02:40,680 --> 00:02:43,040 TALK. 71 00:02:43,040 --> 00:02:43,560 THANK YOU. 72 00:02:43,560 --> 00:02:46,120 >> THANK YOU SO MUCH NIHAL FOR 73 00:02:46,120 --> 00:02:47,800 THE KIND INTRODUCTION AND 74 00:02:47,800 --> 00:02:51,080 INVITATION TO THIS SEMINAR. 75 00:02:51,080 --> 00:02:59,920 I'M COMPUTE EXCITED TO BE HERE I 76 00:02:59,920 --> 00:03:01,320 THINK WE ARE A LITTLE ZOOMED 77 00:03:01,320 --> 00:03:03,120 OUT, THOUGH IT SEEMS IT WILL BE 78 00:03:03,120 --> 00:03:04,680 WITH US FOR QUITE SOME TIME. 79 00:03:04,680 --> 00:03:08,280 AS NIHAL SAID I'M A VIROLOGIST 80 00:03:08,280 --> 00:03:09,200 FROM SAN FRANCISCO, HAVING 81 00:03:09,200 --> 00:03:11,880 WORKED ON MANY VIRUSES IN THE 82 00:03:11,880 --> 00:03:15,200 PAST AND HIV HEPATITIS C AND 83 00:03:15,200 --> 00:03:15,680 ZIKA. 84 00:03:15,680 --> 00:03:18,360 WE HAVE TURNED OUR ATTENTION TO 85 00:03:18,360 --> 00:03:21,240 SARS COV-2 EXACTLY TWO YEARS AGO 86 00:03:21,240 --> 00:03:25,200 WHEN IT BECAME CLEAR THAT THIS 87 00:03:25,200 --> 00:03:27,080 IS GOING TO BE A WORLDWIDE 88 00:03:27,080 --> 00:03:27,680 PROBLEM. 89 00:03:27,680 --> 00:03:28,840 ONE OF THE FIRST THINGS WE HAVE 90 00:03:28,840 --> 00:03:33,760 TO DO WAS THAT WE HAVE TO 91 00:03:33,760 --> 00:03:34,680 PROVIDE THE ENVIRONMENT FOR US 92 00:03:34,680 --> 00:03:37,680 TO STUDY IT AND THAT IS OF 93 00:03:37,680 --> 00:03:40,080 COURSE BSL 3 LABORATORY THAT 94 00:03:40,080 --> 00:03:41,840 ALLOWS RESPIRATORY PATHOGEN 95 00:03:41,840 --> 00:03:42,360 WORK. 96 00:03:42,360 --> 00:03:44,920 WE HAVE HAD -- WE HAVE A LARGE 97 00:03:44,920 --> 00:03:47,080 BSL 3 THAT ALLOWS US TO WORK 98 00:03:47,080 --> 00:03:49,200 WITH BLOOD BORNE PATHOGENS BUT 99 00:03:49,200 --> 00:03:52,000 WE HAVE TO PIVOT AND BUILD A NEW 100 00:03:52,000 --> 00:03:53,240 BSL 3 LABORATORY FOR RESPIRATORY 101 00:03:53,240 --> 00:03:56,440 VIRUSES WHICH WE DID AND 102 00:03:56,440 --> 00:03:59,760 FINISHED ACTUALLY IN EARLY 103 00:03:59,760 --> 00:04:02,400 SUMMER LAST YEAR AND SINCE THEN 104 00:04:02,400 --> 00:04:05,520 WORKING BASICALLY 24/7 IN THAT 105 00:04:05,520 --> 00:04:05,960 LABORATORY. 106 00:04:05,960 --> 00:04:08,200 THIS IS ACTUALLY TAKEN HERE AT 107 00:04:08,200 --> 00:04:10,640 THE UCSF AT ONE OF THE BSL 3 108 00:04:10,640 --> 00:04:12,960 THERE MY COLLEAGUE ANITA, THIS 109 00:04:12,960 --> 00:04:16,080 IS THE FIRST VIRUS THAT WE 110 00:04:16,080 --> 00:04:20,240 CULTURED AND LOOKING FOR 111 00:04:20,240 --> 00:04:21,280 CYTOPATHIC EFFECTS IN CELL 112 00:04:21,280 --> 00:04:22,480 CULTURE HERE. 113 00:04:22,480 --> 00:04:24,560 THE REASON WE BECAME INTERESTED 114 00:04:24,560 --> 00:04:27,440 IN OR WHY WE THOUGHT WE COULD 115 00:04:27,440 --> 00:04:29,080 CONTRIBUTE TO SARS COV-2 116 00:04:29,080 --> 00:04:30,880 RESEARCH WAS BECAUSE WE HAD TWO 117 00:04:30,880 --> 00:04:34,080 ONGOING PROJECTS IN THE LAB. 118 00:04:34,080 --> 00:04:37,280 ONE IS A TESTING PROJECT THAT 119 00:04:37,280 --> 00:04:39,440 NIHAL MENTIONED TOGETHER WITH MY 120 00:04:39,440 --> 00:04:41,640 COLLEAGUE JENNIFER AND DAN 121 00:04:41,640 --> 00:04:44,040 FLETCHER, BOTH FROM BERKELEY BUT 122 00:04:44,040 --> 00:04:46,880 BOTH ALSO HAVE AFFILIATION WITH 123 00:04:46,880 --> 00:04:47,880 GLADSTONE INSTITUTES. 124 00:04:47,880 --> 00:04:50,200 WHERE WE HAD BEEN LOOKING AT 125 00:04:50,200 --> 00:04:53,880 RAPID CRISPER BASED TESTING FOR 126 00:04:53,880 --> 00:04:55,120 HIV. 127 00:04:55,120 --> 00:04:58,360 AND TODAY MORE THAN EVER IT IS 128 00:04:58,360 --> 00:05:00,560 CLEAR THESE TESTS ARE ACTUALLY 129 00:05:00,560 --> 00:05:02,960 EXTREMELY USEFUL FOR SARS COV-2 130 00:05:02,960 --> 00:05:05,200 POTENTIALLY OTHER VIRUSES, WE 131 00:05:05,200 --> 00:05:06,720 REALLY NEED FREQUENT FAST BUT 132 00:05:06,720 --> 00:05:09,960 ALSO IMPORTANTLY ACCURATE TESTS 133 00:05:09,960 --> 00:05:11,000 THAT WE CAN RELY ON. 134 00:05:11,000 --> 00:05:13,880 SO WE PIVOTED PART OF THIS TEST 135 00:05:13,880 --> 00:05:16,000 TO SARS COV-2 AND I WILL TELL 136 00:05:16,000 --> 00:05:18,160 YOU MORE LATER ABOUT THIS. 137 00:05:18,160 --> 00:05:20,600 THE OTHER THING THAT WE HAVE 138 00:05:20,600 --> 00:05:23,080 GOING IN THE LAB THAT WE THOUGHT 139 00:05:23,080 --> 00:05:25,560 COULD BE HELPFUL FOR THE 140 00:05:25,560 --> 00:05:27,280 UNDERSTANDING OF SARS COV-2 IS 141 00:05:27,280 --> 00:05:29,600 WE HAVE BEEN LOOKING TO WORKING 142 00:05:29,600 --> 00:05:31,680 WITH PRIMARY CELL ORGANOID 143 00:05:31,680 --> 00:05:33,840 MODELS ACTUALLY A FEW YEARS BACK 144 00:05:33,840 --> 00:05:36,480 ALREADY, STARTING WITH HEPATITIS 145 00:05:36,480 --> 00:05:38,800 C AND LIVER ORGANOIDS BUT MOVING 146 00:05:38,800 --> 00:05:41,120 TO INTESTINAL ORGANOIDS AND ALSO 147 00:05:41,120 --> 00:05:45,880 INTO HUMAN AIRWAY ORGANOIDS WE 148 00:05:45,880 --> 00:05:46,880 ESTABLISHED FOR SPECIFICALLY 149 00:05:46,880 --> 00:05:48,880 SOME RESEARCH WE WERE DOING ON 150 00:05:48,880 --> 00:05:53,360 INFLUENZA A AND B TOGETHER WITH 151 00:05:53,360 --> 00:05:55,880 VR BY TECHNOLOGY HERE IN SAN 152 00:05:55,880 --> 00:05:57,040 FRANCISCO. 153 00:05:57,040 --> 00:05:58,680 THESE ARE JUST TO MAKE SURE, 154 00:05:58,680 --> 00:06:00,840 THESE ARE ORGANOIDS THAT ARE 155 00:06:00,840 --> 00:06:02,240 ADULT STEM CELL DERIVED SO 156 00:06:02,240 --> 00:06:04,920 COMING DIRECTLY OUT OF BIOPSY OR 157 00:06:04,920 --> 00:06:07,000 RESECTION MATERIAL FROM THE 158 00:06:07,000 --> 00:06:09,360 RESPECTIVE ORGAN. 159 00:06:09,360 --> 00:06:13,520 IT IS METHOD LANCE HAS PIONEER 160 00:06:13,520 --> 00:06:16,160 AD FEW YEARS BACK TO PUT CELLS 161 00:06:16,160 --> 00:06:19,360 IN CULTURE PROLIFERATE THE 162 00:06:19,360 --> 00:06:20,560 PROGENITOR CELLS AND THEN 163 00:06:20,560 --> 00:06:23,280 DIFFERENTIATE THEM INTO THE 164 00:06:23,280 --> 00:06:24,120 INDIVIDUAL ORGANS. 165 00:06:24,120 --> 00:06:27,880 SO HAVING THESE HUMAN AIRWAY 166 00:06:27,880 --> 00:06:30,000 ORGANOIDS AND OTHERS WE THOUGHT 167 00:06:30,000 --> 00:06:32,200 THAT THIS COULD BE IMPORTANT TO 168 00:06:32,200 --> 00:06:33,960 CONTRIBUTE TO SARS COV-2 169 00:06:33,960 --> 00:06:35,760 RESEARCH AT THE TIME. 170 00:06:35,760 --> 00:06:38,080 INDEED, HAVING THE BSL 3, HAVING 171 00:06:38,080 --> 00:06:43,760 THE VIRUSES AND THEN HAVING ALSO 172 00:06:43,760 --> 00:06:46,520 THE INDIVIDUAL ORGAN SYSTEMS 173 00:06:46,520 --> 00:06:49,520 THAT ALLOWED US REALLY TO LOOK 174 00:06:49,520 --> 00:06:51,840 AT SARS COV-2 INFECTION IN 175 00:06:51,840 --> 00:06:54,920 VARIOUS ORGANS, HERE THE LUNG, 176 00:06:54,920 --> 00:06:56,880 INTESTINE BUT ALSO IN THE HEART 177 00:06:56,880 --> 00:06:59,320 CELLS WHERE YOU CAN SEE GREEN OR 178 00:06:59,320 --> 00:07:02,400 HERE IN RED DOUBLE STRANDED RNA 179 00:07:02,400 --> 00:07:03,760 COMPLEX SYSTEM STAINED IN CELLS 180 00:07:03,760 --> 00:07:06,640 AND YOU SEE HOW DRAMATICALLY 181 00:07:06,640 --> 00:07:09,880 BASICALLY THE VIRUS TAKES OVER 182 00:07:09,880 --> 00:07:11,880 THE CELLS AND REPLY CASE IN 183 00:07:11,880 --> 00:07:13,480 THEM, ESPECIALLY THE HEART 184 00:07:13,480 --> 00:07:15,280 CELLS, WE NOTICE TOGETHER WITH 185 00:07:15,280 --> 00:07:20,880 OUR COLLEAGUE COLLEAGUES HERE AT GLADSTONE 186 00:07:20,880 --> 00:07:22,760 THESE CELLS ARE QUIZZICALLY 187 00:07:22,760 --> 00:07:26,200 SENSITIVE TO OXIC EFFECTS OF THE 188 00:07:26,200 --> 00:07:27,960 VIRUS, EVEN ON THROUGH BINDING 189 00:07:27,960 --> 00:07:30,080 BUT ESPECIALLY REPLICATION CELLS 190 00:07:30,080 --> 00:07:35,800 START TO BE EXTREMELY DESTROYED 191 00:07:35,800 --> 00:07:37,360 AND UNDERGOING SOME FORMS OF 192 00:07:37,360 --> 00:07:39,040 CELL DEATH IF THE VIRUS IS 193 00:07:39,040 --> 00:07:39,480 TOUCHING THEM. 194 00:07:39,480 --> 00:07:41,280 SO WE ARE STILL VERY INTERESTED 195 00:07:41,280 --> 00:07:41,760 IN THAT. 196 00:07:41,760 --> 00:07:44,320 WE HAVE COLLABORATED WITH MANY 197 00:07:44,320 --> 00:07:48,800 PEOPLE HERE AT UCSF AND 198 00:07:48,800 --> 00:07:50,680 ELSEWHERE TO LOOK AT THE 199 00:07:50,680 --> 00:07:52,040 PATHOGENESIS OF THE VIRUS IN 200 00:07:52,040 --> 00:07:55,680 THESE DIFFERENT ORGANS. 201 00:07:55,680 --> 00:07:57,800 TODAY I WANT TO TALK ABOUT THREE 202 00:07:57,800 --> 00:08:00,680 SUBJECTS THAT I POINTED OUT IN 203 00:08:00,680 --> 00:08:01,400 MY TITLE. 204 00:08:01,400 --> 00:08:04,480 I THINK WE WANT TO LOOK ABOUT 205 00:08:04,480 --> 00:08:06,760 DETECTION, STUDY AND TREATMENT 206 00:08:06,760 --> 00:08:08,560 AND I WILL DO THIS IN A LITTLE 207 00:08:08,560 --> 00:08:11,520 BIT DIFFERENT ORDER THAN IS 208 00:08:11,520 --> 00:08:13,280 LOGICAL, BUT I WANT TO START 209 00:08:13,280 --> 00:08:15,480 WITH THE HOST DIRECTED THERAPIES 210 00:08:15,480 --> 00:08:16,480 JUST BECAUSE IT IS SOMETHING 211 00:08:16,480 --> 00:08:17,880 THAT WE HAVE A LONG STANDING 212 00:08:17,880 --> 00:08:20,920 INTEREST IN AND WAS ALSO THE 213 00:08:20,920 --> 00:08:22,720 FIRST THINGS WE CONTRIBUTED TO 214 00:08:22,720 --> 00:08:25,800 SARS COV-2. 215 00:08:25,800 --> 00:08:27,360 LET ME START WITH OUR INTEREST 216 00:08:27,360 --> 00:08:29,040 IN HOST DIRECTED THERAPIES. 217 00:08:29,040 --> 00:08:32,080 I THINK IT DEPENDS -- IT REALLY 218 00:08:32,080 --> 00:08:34,960 STEMS FROM THE FACT THAT VIRUSES 219 00:08:34,960 --> 00:08:37,080 ARE UNIQUELY DEPENDENT ON THE 220 00:08:37,080 --> 00:08:42,240 HOST AND THIS DEPENDENCY IS A 221 00:08:42,240 --> 00:08:43,000 ACHILLES HEEL AND OPPORTUNITY 222 00:08:43,000 --> 00:08:45,280 FOR THERAPEUTIC INTERVENTION. 223 00:08:45,280 --> 00:08:47,800 WE HAVE FANTASTIC ANTIVIRALS FOR 224 00:08:47,800 --> 00:08:50,480 HIV AND FOR HEPATITIS C AND 225 00:08:50,480 --> 00:08:54,960 OTHERS BUT ONE OF THE ISSUES 226 00:08:54,960 --> 00:08:57,280 HERE IS THAT THE VIRUS CAN 227 00:08:57,280 --> 00:08:59,480 DEVELOP RAPIDLY DRUG RESISTANCE 228 00:08:59,480 --> 00:09:03,520 AND WE USUALLY HAVE A VERY 229 00:09:03,520 --> 00:09:04,480 HIGHLY SPECIFIC DRUG THAT WORKS 230 00:09:04,480 --> 00:09:06,840 AGAINST MOSTLY ONE PATHOGEN. 231 00:09:06,840 --> 00:09:11,880 IF WE GO INTO THE HOST SIDE THE 232 00:09:11,880 --> 00:09:14,520 IDEA IS THAT WE HAVE A LESS 233 00:09:14,520 --> 00:09:15,760 PROBLEMS WITH DRUG RESISTANCE 234 00:09:15,760 --> 00:09:17,880 BECAUSE WE CANNOT EVOLVE AS FAST 235 00:09:17,880 --> 00:09:19,560 AS THE VIRUS. 236 00:09:19,560 --> 00:09:21,880 AND BECAUSE OF OUR OWN RESEARCH 237 00:09:21,880 --> 00:09:24,440 AND OTHERS WE KNOW THAT MANY 238 00:09:24,440 --> 00:09:28,280 VIRUSES EVEN UNRELATED VIRUSES, 239 00:09:28,280 --> 00:09:31,160 FROM THE CLASS STAND POINT HAVE 240 00:09:31,160 --> 00:09:32,280 DEPENDENCIES ON SIMILAR HOST 241 00:09:32,280 --> 00:09:32,760 PATHWAYS. 242 00:09:32,760 --> 00:09:36,480 SO THE IDEA IS TO FIND THESE 243 00:09:36,480 --> 00:09:38,120 PATHWAYS, TO DEVELOP DRUGS 244 00:09:38,120 --> 00:09:40,040 AGAINST AND WITH THIS TO 245 00:09:40,040 --> 00:09:41,600 ACTUALLY HIT NOT ONLY ONE 246 00:09:41,600 --> 00:09:43,520 PATHOGEN BUT MULTIPLE PATHOGEN, 247 00:09:43,520 --> 00:09:46,400 THE IDEA OF A PAN ANTIVIRAL AND 248 00:09:46,400 --> 00:09:48,880 ALSO THE IDEA OF BEING BETTER 249 00:09:48,880 --> 00:09:50,840 PREPARED FOR SOMETHING THAT SARS 250 00:09:50,840 --> 00:09:53,800 COV-2 THAT IS GOING TO BE 251 00:09:53,800 --> 00:09:55,560 POTENTIALLY EVOLVING IN THE 252 00:09:55,560 --> 00:09:57,880 FUTURE EMERGING VIRUSES THAT WE 253 00:09:57,880 --> 00:10:00,200 HAVE NOT TIED YET TO STUDY IN 254 00:10:00,200 --> 00:10:04,680 DETAIL TO DEVELOP THESE VERY 255 00:10:04,680 --> 00:10:09,080 DETAILED LASER SHARP FOCUS 256 00:10:09,080 --> 00:10:11,280 ANTIVIRALS DIRECT AFFECTING 257 00:10:11,280 --> 00:10:11,960 ANTIVIRALS. 258 00:10:11,960 --> 00:10:13,720 WITH THIS WE HAVE A LONG 259 00:10:13,720 --> 00:10:15,400 STANDING COLLABORATION WITH MY 260 00:10:15,400 --> 00:10:18,920 COLLEAGUE NEVIN KROGAN UCSF AND 261 00:10:18,920 --> 00:10:20,160 SEEN YOUR INVESTIGATOR AT 262 00:10:20,160 --> 00:10:22,800 GLADSTONE AND DIRECTOR OF 263 00:10:22,800 --> 00:10:23,640 QUANTITATIVE BIOLOGY INSTITUTE 264 00:10:23,640 --> 00:10:25,600 HERE AT UCSF WHERE WE HAVE BEEN 265 00:10:25,600 --> 00:10:30,160 LOOKING AT MANY OF THESE MAPS OF 266 00:10:30,160 --> 00:10:33,280 HOST VIRAL INTERACTOMES. 267 00:10:33,280 --> 00:10:35,280 PROTEOMICS BASED MASS 268 00:10:35,280 --> 00:10:36,760 SPECTROMETRY BASED LOOKING AT 269 00:10:36,760 --> 00:10:38,320 EACH INDIVIDUAL VIRAL PROTEINS 270 00:10:38,320 --> 00:10:42,080 AND IDENTIFY BY MASS 271 00:10:42,080 --> 00:10:44,080 SPECTROMETRY THE PROTEINS THAT 272 00:10:44,080 --> 00:10:46,080 CONTACT THESE PROTEINS ARE 273 00:10:46,080 --> 00:10:47,520 INTERACTING WITH VIRAL PROTEINS 274 00:10:47,520 --> 00:10:47,880 IN CELLS. 275 00:10:47,880 --> 00:10:51,360 SO NEVIN AND A LARGE GROUP OF 276 00:10:51,360 --> 00:10:54,280 INVESTIGATORS HERE AT UCSF HAVE 277 00:10:54,280 --> 00:10:56,280 MANAGED TO BUILD UP THE SARS 278 00:10:56,280 --> 00:11:02,360 COV-2 INTERACTOME BY APRIL 2020 279 00:11:02,360 --> 00:11:05,280 WHERE WE COULD IDENTIFY 332 SARS 280 00:11:05,280 --> 00:11:07,200 COV-2 INTERACTERS AND BY JUST 281 00:11:07,200 --> 00:11:10,040 LOOKING INTO THESE INTERACTERS, 282 00:11:10,040 --> 00:11:13,280 69 OF THEM WERE DRUGGABLE HOST 283 00:11:13,280 --> 00:11:14,760 CHARACTERS WHERE FDA APPROVED 284 00:11:14,760 --> 00:11:17,400 DRUGS WERE AVAILABLE AND SO THIS 285 00:11:17,400 --> 00:11:20,440 WAS THE FIRST START TO LOOK INTO 286 00:11:20,440 --> 00:11:22,720 THE POTENTIAL OF HOST DIRECTED 287 00:11:22,720 --> 00:11:24,640 THERAPIES AGAINST SARS COV-2 AND 288 00:11:24,640 --> 00:11:27,520 THIS WORK IS VERY MUCH ONGOING 289 00:11:27,520 --> 00:11:29,760 WITH MANY BASIC BIOLOGISTS HERE 290 00:11:29,760 --> 00:11:32,080 AT UCSF LOOKING INTO 291 00:11:32,080 --> 00:11:33,320 STRUCTURALLY OR FUNCTIONALLY 292 00:11:33,320 --> 00:11:39,680 INTO THESE INTERACTIONS AND US 293 00:11:39,680 --> 00:11:40,720 LIKE OTHER VIROLOGISTS AROUND 294 00:11:40,720 --> 00:11:41,880 THE WORLD TESTING WHETHER ANY OF 295 00:11:41,880 --> 00:11:44,240 THESE DRUGS HAVE AN EFFECT ON 296 00:11:44,240 --> 00:11:48,480 SARS COV-2 IN VITRO OR IN MOUSE 297 00:11:48,480 --> 00:11:50,080 OR HAMSTER MODELS. 298 00:11:50,080 --> 00:11:52,280 THE OTHER WAY HOW YOU CAN 299 00:11:52,280 --> 00:11:53,720 APPROACH HOST DIRECTED THERAPIES 300 00:11:53,720 --> 00:11:55,960 IS NOT JUST LOOKING AT THIS 301 00:11:55,960 --> 00:11:57,480 PROTEINS THAT ARE INTERACTING 302 00:11:57,480 --> 00:11:59,920 WITH VIRAL PROTEINS, YOU CAN 303 00:11:59,920 --> 00:12:02,280 ACTUALLY LOOK FUNCTIONALLY AT 304 00:12:02,280 --> 00:12:03,960 THE WHOLE -- AT ALL HUMAN 305 00:12:03,960 --> 00:12:05,680 PROTEINS AND SEE WHICH ONES OF 306 00:12:05,680 --> 00:12:07,720 THEM ARE ACTUALLY DEPENDENCY 307 00:12:07,720 --> 00:12:10,280 FACTORS OF THE VIRUS, THIS IS A 308 00:12:10,280 --> 00:12:12,640 CLASSICAL CRISPR SCREEN WHERE 309 00:12:12,640 --> 00:12:16,800 YOU KNOCK OUT INDIVIDUAL HOST 310 00:12:16,800 --> 00:12:18,120 PROTEINS, INFECT THE CELL AND 311 00:12:18,120 --> 00:12:20,600 THEN IN THIS CASE IT IS A LIVE 312 00:12:20,600 --> 00:12:21,600 DEATH SELECTION AND LOOK FOR 313 00:12:21,600 --> 00:12:22,600 SURVIVING CELLS THAT ARE NOT 314 00:12:22,600 --> 00:12:26,280 KILLED BY THE VIRAL REPLICATION. 315 00:12:26,280 --> 00:12:27,920 SEQUENCE THE GENE THAT IS 316 00:12:27,920 --> 00:12:29,600 EXPRESSED OR THAT IS KNOCKED 317 00:12:29,600 --> 00:12:33,560 DOWN IN THE CELLS AND THEN 318 00:12:33,560 --> 00:12:35,800 ASSEMBLE THE LIST OF THE 319 00:12:35,800 --> 00:12:38,680 DEPENDENCY FACTORS FOR AN 320 00:12:38,680 --> 00:12:39,280 INDIVIDUAL VIRUS. 321 00:12:39,280 --> 00:12:42,120 SO WE DID THIS ALSO VERY EARLY 322 00:12:42,120 --> 00:12:44,600 IN 2020 TOGETHER WITH MY 323 00:12:44,600 --> 00:12:48,280 COLLEAGUE ANDREAIAS PUSHNIK AT 324 00:12:48,280 --> 00:12:51,920 THE ZUCKERBERG BIOHUB AT SAN 325 00:12:51,920 --> 00:12:54,360 FRANCISCO AND WE ARE NOT ONLY 326 00:12:54,360 --> 00:12:56,280 FOCUSED ON SARS COV-2 BUT 327 00:12:56,280 --> 00:12:58,560 PARALLEL INCLUDE TWO COMMON COLD 328 00:12:58,560 --> 00:13:02,640 CORONA VIRUSES 229E AND OC 43 329 00:13:02,640 --> 00:13:05,640 JUST TO GO PRECISELY AT THE 330 00:13:05,640 --> 00:13:09,160 QUESTION ARE THEY PAN VIRAL 331 00:13:09,160 --> 00:13:10,080 FACTORS THAT WE CAN TARGET IN 332 00:13:10,080 --> 00:13:14,680 THE FUTURE TO NOT ONLY TARGET OR 333 00:13:14,680 --> 00:13:18,440 HIT SARS COV-2 BUT ALSO RELATED 334 00:13:18,440 --> 00:13:19,120 CORONA VIRUSES. 335 00:13:19,120 --> 00:13:21,680 THIS IS THE WORK THAT WAS DONE 336 00:13:21,680 --> 00:13:26,640 BY PETER WONG AND ANDREIAS'S LAB 337 00:13:26,640 --> 00:13:28,680 AND MY LAB, CAMILLE HAS SINCE 338 00:13:28,680 --> 00:13:30,280 GRADUATED AND IS POST-DOC IN THE 339 00:13:30,280 --> 00:13:33,040 LAB READY TO TAKE HER NEXT STEP. 340 00:13:33,040 --> 00:13:36,120 SO WHEN WE DID THIS AS I 341 00:13:36,120 --> 00:13:38,600 EXPLAINED BEFORE, WE IDENTIFIED 342 00:13:38,600 --> 00:13:40,840 SEVERAL HOST FACTORS FOR SARS 343 00:13:40,840 --> 00:13:44,280 COV-2 ENRICHED IN THIS SCREEN, 344 00:13:44,280 --> 00:13:48,720 VERY IMPORTANTLY WE IDENTIFIED H 345 00:13:48,720 --> 00:13:50,480 2 AS THE ENTRY FACTOR BUT 346 00:13:50,480 --> 00:13:52,720 NOTICED THERE WERE QUITE A FEW 347 00:13:52,720 --> 00:13:55,480 PROTEINS THAT ARE IMPORTANT IN 348 00:13:55,480 --> 00:13:58,880 CHOLESTEROL HOMEOSTASIS AS WELL 349 00:13:58,880 --> 00:14:02,200 AS LYSOSOME AUTOPHAGESOME 350 00:14:02,200 --> 00:14:04,240 RELATED FUNCTIONS. 351 00:14:04,240 --> 00:14:07,480 229E HAD MORE PROTEINS THAN WE 352 00:14:07,480 --> 00:14:08,160 IDENTIFIED. 353 00:14:08,160 --> 00:14:10,840 AGAIN, THE RECEPTOR WAS AMONG 354 00:14:10,840 --> 00:14:14,600 THE HIGHEST ENRICHED GENES BUT 355 00:14:14,600 --> 00:14:16,680 WE ALSO FOUND QUITE A FEW GOLGI 356 00:14:16,680 --> 00:14:18,440 RELATED FUNCTION PROTEINS THAT 357 00:14:18,440 --> 00:14:20,240 WERE HIGHLY ENRICHED. 358 00:14:20,240 --> 00:14:24,920 WHILE THE OC 43 IS A GLYCAN 359 00:14:24,920 --> 00:14:26,920 BASED RECEPTOR AND ENTERING 360 00:14:26,920 --> 00:14:30,280 VIRUS SO THE GLYCOSAMINOGLYCAN 361 00:14:30,280 --> 00:14:31,840 BIOSYNTHESIS PATHWAY WAS HIGHLY 362 00:14:31,840 --> 00:14:34,640 ENRICHED IN THESE AMONG OTHERS 363 00:14:34,640 --> 00:14:36,280 THAT WERE SHARED WITH THE OTHER 364 00:14:36,280 --> 00:14:37,560 TWO CORONA VIRUSES. 365 00:14:37,560 --> 00:14:40,480 WHEN WE LOOK AT THE OVERLAP 366 00:14:40,480 --> 00:14:42,560 BETWEEN THESE THREE VIRUSES IN 367 00:14:42,560 --> 00:14:46,200 TERMS OF INDIVIDUAL GENES, 368 00:14:46,200 --> 00:14:48,080 COMPARING HERE SARS COV-2 AND 369 00:14:48,080 --> 00:14:51,080 229E WE DIDN'T SEE MUCH OVERLAP, 370 00:14:51,080 --> 00:14:54,200 JUST THE LDL RECEPTOR HERE. 371 00:14:54,200 --> 00:14:55,520 LITTLE BIT MORE BETWEEN SARS 372 00:14:55,520 --> 00:14:58,440 COV-2 AND OC 43. 373 00:14:58,440 --> 00:15:00,200 BUT THEN QUITE A BIT BETWEEN THE 374 00:15:00,200 --> 00:15:02,880 TWO COMMON COLD CORONA VIRUSES. 375 00:15:02,880 --> 00:15:04,600 SO THIS EITHER INDICATED THAT 376 00:15:04,600 --> 00:15:07,040 SARS COV-2 WAS UNIQUE, DIFFERENT 377 00:15:07,040 --> 00:15:09,560 FROM THEM, BUT WE ALSO THOUGHT 378 00:15:09,560 --> 00:15:11,880 IT MIGHT BE THAT AT -- THAT THE 379 00:15:11,880 --> 00:15:13,320 OVERLAP MIGHT NOT BE HAPPENING 380 00:15:13,320 --> 00:15:16,080 AT THE GENE LEVEL BUT MIGHT BE 381 00:15:16,080 --> 00:15:19,320 HAPPENING AT THE PATHWAY LEVEL. 382 00:15:19,320 --> 00:15:23,080 SO WE WORKED WITH NEVIN'S GROUP 383 00:15:23,080 --> 00:15:25,880 AND DID A NETWORK PROPAGATION 384 00:15:25,880 --> 00:15:29,920 ANALYSIS WHERE WE REALLY LOOKED 385 00:15:29,920 --> 00:15:32,160 AT THE NETWORKS THAT WERE 386 00:15:32,160 --> 00:15:35,920 ENRICHED BETWEEN THE THREE VIRAL 387 00:15:35,920 --> 00:15:36,400 INFECTIONS. 388 00:15:36,400 --> 00:15:38,440 WE COULD IDENTIFY QUITE A FEW, 389 00:15:38,440 --> 00:15:40,440 ONE WAS CHOLESTEROL METABOLIC 390 00:15:40,440 --> 00:15:42,080 PATHWAY, ALSO IT WAS HIGHLY 391 00:15:42,080 --> 00:15:44,200 ENRICHED WITH SARS COV-2, THE 392 00:15:44,200 --> 00:15:47,760 OTHER PROTEINS, THE OTHER 393 00:15:47,760 --> 00:15:50,560 VIRUSES ALSO HAD CHOLESTEROL 394 00:15:50,560 --> 00:15:52,480 RELATED CO-FACTORS THAT WERE 395 00:15:52,480 --> 00:15:55,760 ABSOLUTELY CRITICAL FOR THEIR 396 00:15:55,760 --> 00:15:56,680 REPLICATION. 397 00:15:56,680 --> 00:15:59,080 THE SAME, ANOTHER PATHWAY WE 398 00:15:59,080 --> 00:16:01,640 IDENTIFIED WAS MACRO AUTOPHAGY 399 00:16:01,640 --> 00:16:06,320 AND PHOSPHOMETABOLISM TYROSINE 400 00:16:06,320 --> 00:16:09,640 KINASE, HIGHLY ENRICHED BETWEEN 401 00:16:09,640 --> 00:16:11,960 THE THREE PROTEIN WITH DIFFERENT 402 00:16:11,960 --> 00:16:14,080 INDIVIDUAL FACTORS. 403 00:16:14,080 --> 00:16:17,920 TO START TO LOOK INTO VALIDATION 404 00:16:17,920 --> 00:16:19,160 HERE, WE DID INDIVIDUAL KNOCK 405 00:16:19,160 --> 00:16:24,000 OUT EXPERIMENTS WITH CRISPR 406 00:16:24,000 --> 00:16:26,080 HERE, FOR EXAMPLE WE KNOCKED 407 00:16:26,080 --> 00:16:28,560 DOWN THE SARS COV-2 HITS, 408 00:16:28,560 --> 00:16:30,120 SPECIFICALLY THE HITS WE 409 00:16:30,120 --> 00:16:32,680 IDENTIFIED IN THE CHOLESTEROL 410 00:16:32,680 --> 00:16:34,640 PATHWAY BUT ALSO OTHER RELATED 411 00:16:34,640 --> 00:16:35,760 FACTORS AND YOU CAN SEE NICELY 412 00:16:35,760 --> 00:16:37,560 THAT IF YOU KNOCK IT DOWN AND 413 00:16:37,560 --> 00:16:39,600 YOU INFECT YOU SEE THAT YOU KILL 414 00:16:39,600 --> 00:16:42,120 INFECTION, IF YOU REPLACE THE 415 00:16:42,120 --> 00:16:43,800 FACTOR CLASSICAL 416 00:16:43,800 --> 00:16:44,440 RESUPPLEMENTATION EXPERIMENTS 417 00:16:44,440 --> 00:16:46,800 YOU CAN SEE THAT THIS IS 418 00:16:46,800 --> 00:16:49,640 SPECIFIC BECAUSE YOU ENABLE 419 00:16:49,640 --> 00:16:50,480 INFECTION AGAIN. 420 00:16:50,480 --> 00:16:52,240 THIS WAS SHOWN WITH MULTIPLE 421 00:16:52,240 --> 00:16:54,440 FACTORS AND YOU CAN SEE THAT'S 422 00:16:54,440 --> 00:16:56,760 SPECIALLY ALSO THESE CHOLESTEROL 423 00:16:56,760 --> 00:16:58,880 PATHWAY FACTORS WERE HIGHLY 424 00:16:58,880 --> 00:17:00,160 IMPORTANT FOR THE REPLICATION OF 425 00:17:00,160 --> 00:17:01,200 THE VIRUS. 426 00:17:01,200 --> 00:17:03,880 WHEN WE DID THE SAME THING WITH 427 00:17:03,880 --> 00:17:05,600 THE TWO OTHER COMMON COLD CORONA 428 00:17:05,600 --> 00:17:09,320 VIRUSES WE SAW THAT ALSO NOT ALL 429 00:17:09,320 --> 00:17:12,520 BUT MANY ESPECIALLY THE 430 00:17:12,520 --> 00:17:15,720 CHOLESTEROL PATHWAY, FACTORS 431 00:17:15,720 --> 00:17:17,960 WERE ALSO IMPORTANT FOR ENABLING 432 00:17:17,960 --> 00:17:19,720 VIRAL REPLICATION FOR THE COMMON 433 00:17:19,720 --> 00:17:21,400 COLD CORONA VIRUSES, REALLY 434 00:17:21,400 --> 00:17:22,720 UNDERSCORING THE VALIDITY OF 435 00:17:22,720 --> 00:17:24,080 THIS NETWORK PROPAGATION 436 00:17:24,080 --> 00:17:25,480 ANALYSIS AND THE IDENTIFICATION 437 00:17:25,480 --> 00:17:30,320 OF THE CHOLESTEROL PATHWAY AS 438 00:17:30,320 --> 00:17:32,080 POTENTIAL PAN VIRAL PATHWAY FOR 439 00:17:32,080 --> 00:17:33,480 CORONA VIRUSES. 440 00:17:33,480 --> 00:17:35,720 WE DID THIS ALSO WITH OTHER 441 00:17:35,720 --> 00:17:39,600 VIRUSES HERE WITH OC 43, KNOCK 442 00:17:39,600 --> 00:17:42,600 DOWN IMPORTANT FACTORS AND 443 00:17:42,600 --> 00:17:44,800 ENDOSOME MATURATION AND PI 3 444 00:17:44,800 --> 00:17:46,920 KINASE COMPLEX HIGHLY ENRICHED, 445 00:17:46,920 --> 00:17:48,680 YOU CAN SEE REALLY IMPORTANT FOR 446 00:17:48,680 --> 00:17:49,280 REPLICATION. 447 00:17:49,280 --> 00:17:50,720 THEY WERE ALSO IMPORTANT FOR 448 00:17:50,720 --> 00:17:53,360 REPLICATION FOR 229E ALSO TO 449 00:17:53,360 --> 00:17:54,960 DIFFERENT DEGREE AND WHEN WE DID 450 00:17:54,960 --> 00:17:58,160 THIS WITH SARS COV-2 YOU CAN SEE 451 00:17:58,160 --> 00:18:00,280 THAT ALL BUT TWO WERE IMPORTANT 452 00:18:00,280 --> 00:18:02,480 FOR SARS COV-2 REPLICATION 453 00:18:02,480 --> 00:18:04,480 ESPECIALLY ALSO IN THE PI 3 454 00:18:04,480 --> 00:18:05,280 KINASE COMPLEX. 455 00:18:05,280 --> 00:18:07,480 SO THESE ARE -- THIS SHOWS YOU 456 00:18:07,480 --> 00:18:12,080 THE POWER OF THE METHOD BUT ALSO 457 00:18:12,080 --> 00:18:13,400 THE OPPORTUNITIES AND REALLY 458 00:18:13,400 --> 00:18:15,080 DIGGING DEEPER AND UNDERSTANDING 459 00:18:15,080 --> 00:18:16,000 THE INDIVIDUAL LIFE CYCLE OF 460 00:18:16,000 --> 00:18:18,960 THESE VIRUSES, BUT ALSO 461 00:18:18,960 --> 00:18:21,200 IDENTIFYING POTENTIALLY COMMON 462 00:18:21,200 --> 00:18:22,240 TARGETS THAT MIGHT BE IMPORTANT 463 00:18:22,240 --> 00:18:23,840 FOR THERAPEUTICS. 464 00:18:23,840 --> 00:18:26,080 SO THE QUESTION NOW WAS IS THIS 465 00:18:26,080 --> 00:18:27,920 REALLY SOMETHING THAT WE CAN 466 00:18:27,920 --> 00:18:29,280 DERIVE FROM THIS DATA. 467 00:18:29,280 --> 00:18:31,920 CAN WE IDENTIFY PAN CORONA VIRUS 468 00:18:31,920 --> 00:18:33,720 PHARMACOLOGICAL INHIBITORS. 469 00:18:33,720 --> 00:18:36,440 SO WE FOCUSED ON PI 3 KINASE 470 00:18:36,440 --> 00:18:38,960 INHIBITORS HERE. 471 00:18:38,960 --> 00:18:41,880 WHERE WE LOOK FOR INFECTION IN 472 00:18:41,880 --> 00:18:45,720 SARS COV-2, 229E AND OC 43 YOU 473 00:18:45,720 --> 00:18:48,760 CAN SEE THIS INHIBITOR ALSO 474 00:18:48,760 --> 00:18:52,200 REALLY HAS VERY PRONOUNCED 475 00:18:52,200 --> 00:18:53,520 INHIBITORY ACTIVITY FOR ALL 476 00:18:53,520 --> 00:18:57,360 THREE FACTORS, WHILE NOT BEING 477 00:18:57,360 --> 00:18:58,080 TOXIC. 478 00:18:58,080 --> 00:19:03,040 THE SAME IS FOR CHOLESTEROL 479 00:19:03,040 --> 00:19:04,040 INTRACELLULAR CHOLESTEROL 480 00:19:04,040 --> 00:19:05,960 PRODUCTION INHIBITOR HERE, WHERE 481 00:19:05,960 --> 00:19:08,680 YOU CAN SEE ESPECIALLY THE LOWER 482 00:19:08,680 --> 00:19:12,680 DOSE YOU SEE ALSO AN INTERESTING 483 00:19:12,680 --> 00:19:14,480 INHIBITION OF THESE TWO VIRUSES, 484 00:19:14,480 --> 00:19:18,240 A LITTLE LESS WITH OC 43 AND 485 00:19:18,240 --> 00:19:19,320 HIGHER CONCENTRATION HERE WE 486 00:19:19,320 --> 00:19:22,720 HAVE MORE TOXICITY BUT THIS IS 487 00:19:22,720 --> 00:19:24,480 TYPE OF STUDIES WE HAVE BEEN 488 00:19:24,480 --> 00:19:26,280 DOING WITH OTHER VIRUSES BEFORE 489 00:19:26,280 --> 00:19:27,880 AND WE CONTINUE TO DO WITH THIS 490 00:19:27,880 --> 00:19:30,280 VIRUS HERE AND THAT WE BRING NOW 491 00:19:30,280 --> 00:19:33,280 INTO IN VIVO STUDIES TO SEE 492 00:19:33,280 --> 00:19:35,640 REALLY POTENTIAL AND ALSO THE 493 00:19:35,640 --> 00:19:39,480 PRECISE MOLECULAR FUNCTION OF 494 00:19:39,480 --> 00:19:41,680 THESE IN THE LIFE CYCLE. 495 00:19:41,680 --> 00:19:43,840 IF I SUMMARIZE THIS FIRST PART I 496 00:19:43,840 --> 00:19:47,240 THINK THE CRISPR KNOCK OUT 497 00:19:47,240 --> 00:19:48,600 SCREENS HAVE BEEN BY US AND 498 00:19:48,600 --> 00:19:50,960 OTHERS PERFORMED MANY OTHERS 499 00:19:50,960 --> 00:19:52,800 HAVE DONE SIMILAR SCREENS WITH 500 00:19:52,800 --> 00:19:55,240 HOST FACTORS CRITICAL FOR SARS 501 00:19:55,240 --> 00:19:56,840 COV-2 BUT FOR US ALSO WE 502 00:19:56,840 --> 00:19:59,800 COMPARED WITH OC 43 AND 229E 503 00:19:59,800 --> 00:20:02,400 INFECTION AND THAT HELPED US TO 504 00:20:02,400 --> 00:20:03,720 IDENTIFY SHARED ESSENTIAL 505 00:20:03,720 --> 00:20:05,840 PATHWAYS BETWEEN THE VIRUSES 506 00:20:05,840 --> 00:20:08,440 INCLUDING CHOLESTEROL SYNTHESIS 507 00:20:08,440 --> 00:20:11,040 AND PHOSPHOKINASE, KINASE 508 00:20:11,040 --> 00:20:12,960 PATHWAYS WERE IDENTIFIED AND WE 509 00:20:12,960 --> 00:20:15,080 ARE CONTINUING TO WORK ON IT 510 00:20:15,080 --> 00:20:16,120 NIHAL MENTIONED WE HAVE A LONG 511 00:20:16,120 --> 00:20:17,320 STANDING INTEREST IN LIPIDS AND 512 00:20:17,320 --> 00:20:19,360 VIRAL INFECTION STEMMING FROM 513 00:20:19,360 --> 00:20:22,320 OUR WORK IN HEPATITIS C AND WE 514 00:20:22,320 --> 00:20:24,280 ARE VERY INTERESTED IN FOLLOWING 515 00:20:24,280 --> 00:20:26,080 UP WITH CHOLESTEROL SYNTHESIS 516 00:20:26,080 --> 00:20:28,960 PATHWAY, I THINK IT IS CLEAR 517 00:20:28,960 --> 00:20:32,080 THAT ENVELOPE VIRUSES HAVE A 518 00:20:32,080 --> 00:20:34,680 UNIQUE AFFINITY TO CHOLESTEROL 519 00:20:34,680 --> 00:20:35,920 SYNTHESIS BUT WE THINK THERE 520 00:20:35,920 --> 00:20:39,240 MIGHT BE MORE TO THE CHOLESTEROL 521 00:20:39,240 --> 00:20:41,560 CONNECTION WITH SARS COV-2 ALSO 522 00:20:41,560 --> 00:20:43,880 GIVEN OBVIOUS RISK OF OBESITY AS 523 00:20:43,880 --> 00:20:47,520 A RISK FACTOR FOR SEVERE 524 00:20:47,520 --> 00:20:48,120 DISEASE. 525 00:20:48,120 --> 00:20:51,520 SO WE ARE OVERALL INTERESTING TO 526 00:20:51,520 --> 00:20:53,520 EITHER USE EXISTING 527 00:20:53,520 --> 00:20:55,040 PHARMACOLOGICAL INHIBITORS OR 528 00:20:55,040 --> 00:20:56,560 DERIVE NEW INHIBITORS OF 529 00:20:56,560 --> 00:21:00,680 IDENTIFIED PATHWAYS TO RESTRICT 530 00:21:00,680 --> 00:21:02,840 PAN CORONA VIRUS REPLICATION TO 531 00:21:02,840 --> 00:21:05,480 GET AHEAD OF VARIANTS BUT ALSO 532 00:21:05,480 --> 00:21:06,640 NEW AND EMERGING VIRUSES. 533 00:21:06,640 --> 00:21:08,840 SO I THINK THIS IS A WHOLE NEW 534 00:21:08,840 --> 00:21:10,600 FIELD, I THINK THE PHARMA 535 00:21:10,600 --> 00:21:12,640 COMPANIES ARE MUCH MORE OPEN TO 536 00:21:12,640 --> 00:21:15,160 THIS IDEA BEFORE THEY WERE NOT 537 00:21:15,160 --> 00:21:17,400 BUT I THINK SARS COV-2 HAS 538 00:21:17,400 --> 00:21:19,680 CHANGED MANY PERSPECTIVES HERE 539 00:21:19,680 --> 00:21:22,480 AND I THINK THAT HOST DIRECTED 540 00:21:22,480 --> 00:21:24,720 THERAPY COULD BE A VERY GOOD 541 00:21:24,720 --> 00:21:26,440 ADDITION TO COMBINATION THERAPY 542 00:21:26,440 --> 00:21:28,480 OF ANTIVIRALS FOR SARS COV-2 IN 543 00:21:28,480 --> 00:21:31,280 THE FUTURE GIVEN IT WILL REDUCE 544 00:21:31,280 --> 00:21:33,200 RESISTANCE DEVELOPMENT. 545 00:21:33,200 --> 00:21:35,080 NOW I SHIFT TO THE SECOND PART 546 00:21:35,080 --> 00:21:36,280 OF MY TALK WHICH IS THE 547 00:21:36,280 --> 00:21:36,640 DETECTION PART. 548 00:21:36,640 --> 00:21:40,400 AND THE QUESTION HOW CAN WE GET 549 00:21:40,400 --> 00:21:44,360 TO BETTER SARS COV-2 TESTS. 550 00:21:44,360 --> 00:21:45,800 WE ALL PROBABLY HAVE ANTIGEN 551 00:21:45,800 --> 00:21:47,680 TESTS NOW AT HOME TESTING OUR 552 00:21:47,680 --> 00:21:49,000 KIDS, TESTING OURSELVES WHEN WE 553 00:21:49,000 --> 00:21:52,960 WANT TO GET INTO THE COUNTRY, 554 00:21:52,960 --> 00:21:55,280 HAVING SENTINEL TESTING IN 555 00:21:55,280 --> 00:21:57,840 INSTITUTES, BUT THE QUESTION IS 556 00:21:57,840 --> 00:21:59,160 ALWAYS THEY ARE FREQUENT AND 557 00:21:59,160 --> 00:22:03,120 FAST AND CHEAP BUT ARE THEY ALSO 558 00:22:03,120 --> 00:22:03,880 ACCURATE. 559 00:22:03,880 --> 00:22:06,360 SO AS I SAID WE HAVE BEEN 560 00:22:06,360 --> 00:22:07,800 THINKING ABOUT THESE TYPES OF 561 00:22:07,800 --> 00:22:11,120 TESTS FOR QUITE A WHILE. 562 00:22:11,120 --> 00:22:15,600 THIS IS MY -- MY COLLEAGUE DAN 563 00:22:15,600 --> 00:22:17,800 FLETCHER AND JENNIFER DOUDNA 564 00:22:17,800 --> 00:22:19,680 FROM BERKELEY BUT HAVE 565 00:22:19,680 --> 00:22:24,080 AFFILIATION AT GLADSTONE. 566 00:22:24,080 --> 00:22:26,880 AND WE REALLY SET OUT TO COMBINE 567 00:22:26,880 --> 00:22:29,840 OUR RESPECTIVE EXPERTISE IN 568 00:22:29,840 --> 00:22:32,280 VIROLOGY CRISPR TECHNOLOGY AND 569 00:22:32,280 --> 00:22:34,800 DAN WHO IS A BIOENGINEER AND HAS 570 00:22:34,800 --> 00:22:36,280 PIONEERED THE WORK IN USING 571 00:22:36,280 --> 00:22:40,720 MOBILE PHONES AS DIAGNOSTICS. 572 00:22:40,720 --> 00:22:43,080 AT THE CENTER OF OUR TECHNOLOGY 573 00:22:43,080 --> 00:22:46,600 IS THE ENZYME CALLED CAS 13A, IT 574 00:22:46,600 --> 00:22:50,240 IS A -- ONE OF THE CRISPR 575 00:22:50,240 --> 00:22:52,000 ENZYMES, IT IS AN RNA BINDING 576 00:22:52,000 --> 00:22:55,640 AND EDITING CRISPR ENZYME. 577 00:22:55,640 --> 00:22:58,160 SO IF YOU COMBINE IT WITH 578 00:22:58,160 --> 00:23:00,280 APPROPRIATE GUIDE RNA HERE FOR 579 00:23:00,280 --> 00:23:04,960 SARS COV-2 YOU TARGET BASICALLY 580 00:23:04,960 --> 00:23:07,760 THE ENZYME TO THE TARGET, IT 581 00:23:07,760 --> 00:23:09,240 BECOMES ACTIVE AND IT STARTS 582 00:23:09,240 --> 00:23:11,760 EDITING BECAUSE THAT'S WHAT IT 583 00:23:11,760 --> 00:23:13,880 DOES, US BASICALLY IS A NUCLEUS 584 00:23:13,880 --> 00:23:17,880 AND THE INTERESTING PART ABOUT 585 00:23:17,880 --> 00:23:19,880 CAS 13 IS IT IS NOT A DIRECTED 586 00:23:19,880 --> 00:23:23,120 EDITING BUT IT ALSO EDITS ANY 587 00:23:23,120 --> 00:23:24,480 RNA THAT IS COLLATERALLY 588 00:23:24,480 --> 00:23:26,840 SUPPLIED TO THE REACTION, SO YOU 589 00:23:26,840 --> 00:23:30,680 CAN USE THIS BY JUST GIVING IT A 590 00:23:30,680 --> 00:23:36,080 REPORTER RNA WITH A QUENCHER AND 591 00:23:36,080 --> 00:23:37,880 FLOR FORE, IF THIS RNA IS 592 00:23:37,880 --> 00:23:40,520 CLEAVED THE QUENCHER IS RELEASED 593 00:23:40,520 --> 00:23:42,280 AND FLUORESCENCE IS STARTING TO 594 00:23:42,280 --> 00:23:46,680 GLOW AND IT CAN BE CAPTURED WITH 595 00:23:46,680 --> 00:23:48,480 FOR EXAMPLE A MOBILE PHONE OR 596 00:23:48,480 --> 00:23:53,680 WITH A CLASSICAL PLATE READER IN 597 00:23:53,680 --> 00:23:55,680 THE LAB. 598 00:23:55,680 --> 00:23:57,320 SO INITIALLY -- SO THIS IS A 599 00:23:57,320 --> 00:23:58,920 VERY SIMPLE REACTION. 600 00:23:58,920 --> 00:24:04,360 IT IS BASICALLY AN ANTIGEN TEST 601 00:24:04,360 --> 00:24:07,000 FOR RNA, NOT OF COURSE WITH 602 00:24:07,000 --> 00:24:09,680 ANTIBODIES USUALLY BUT IT IS A 603 00:24:09,680 --> 00:24:12,440 DIRECT TARGETING OF THE -- OF 604 00:24:12,440 --> 00:24:15,360 DETECTING AGENT TO VIRAL RNA. 605 00:24:15,360 --> 00:24:20,840 THIS ALLOWS US TO POTENTIALLY BE 606 00:24:20,840 --> 00:24:24,440 AS CONVENIENT AS ANTIGEN TEST 607 00:24:24,440 --> 00:24:26,240 BUT MORE PRECISE OR AS PRECISE 608 00:24:26,240 --> 00:24:27,680 AS MOLECULAR TEST, THAT WAS THE 609 00:24:27,680 --> 00:24:32,840 GOAL WHEN WE STARTED OUT TO MAKE 610 00:24:32,840 --> 00:24:34,400 IT PORTABLE WE SWITCHED TO THE 611 00:24:34,400 --> 00:24:39,280 MOBILE PHONE AS DETECTOR. 612 00:24:39,280 --> 00:24:41,480 BUT MANY PEOPLE DOUBTED THIS 613 00:24:41,480 --> 00:24:44,960 SIMPLE REACTION WOULD BE 614 00:24:44,960 --> 00:24:47,440 SENSITIVE ENOUGH BECAUSE IT 615 00:24:47,440 --> 00:24:49,960 DOESN'T HAVE -- THE INITIAL DATA 616 00:24:49,960 --> 00:24:53,360 WERE NOT PROMISING WHEN YOU DID 617 00:24:53,360 --> 00:24:55,560 NOT TWEAK THE ESSAY AS I'M 618 00:24:55,560 --> 00:24:58,560 SHOWING WE HAVE DONE SINCE BUT 619 00:24:58,560 --> 00:25:01,920 ALSO MOST OF THE CRISPR 620 00:25:01,920 --> 00:25:03,880 REACTIONS YOU SEE TODAY ARE ANY 621 00:25:03,880 --> 00:25:05,600 COMMERCIAL CRISPR OR 622 00:25:05,600 --> 00:25:08,040 COMMERCIALLY DEVELOPED CRISPR 623 00:25:08,040 --> 00:25:09,360 REACTIONS INCLUDING 624 00:25:09,360 --> 00:25:11,680 AMPLIFICATION STEP IN THEIR 625 00:25:11,680 --> 00:25:12,480 REACTION. 626 00:25:12,480 --> 00:25:14,440 WHERE REVERSE TRANSCRIBING THE 627 00:25:14,440 --> 00:25:17,560 INITIAL RNA AMPLIFYING AND FOR 628 00:25:17,560 --> 00:25:19,720 TRANSCRIBING BEGIN TO MAKE 629 00:25:19,720 --> 00:25:20,120 CRISPR TEST. 630 00:25:20,120 --> 00:25:21,480 WHEN WE SET OUT TO DEVELOP THIS 631 00:25:21,480 --> 00:25:23,880 TEST WE REALLY PURPOSEFULLY 632 00:25:23,880 --> 00:25:27,600 DECIDEDECIDED THAT WE WEREN'T REALLY 633 00:25:27,600 --> 00:25:29,720 PUSH THE ENVELOPE ON THIS DIRECT 634 00:25:29,720 --> 00:25:31,920 DETECTION ESSAY WHERE WE DO NOT 635 00:25:31,920 --> 00:25:33,600 INCLUDE ANY AMPLIFICATION STEP. 636 00:25:33,600 --> 00:25:36,560 WHEN WE STARTED TO SHIFT ESSAY 637 00:25:36,560 --> 00:25:38,480 FROM HIV TO SARS COV-2 IT WAS 638 00:25:38,480 --> 00:25:40,280 LAST YEAR WHEN SEQUENCE OF SARS 639 00:25:40,280 --> 00:25:42,240 TWO YEARS AGO WHEN THE -- WHEN 640 00:25:42,240 --> 00:25:43,880 SARS -- WHEN THE SARS COV-2 641 00:25:43,880 --> 00:25:47,080 SEQUENCE BECAME AVAILABLE WE 642 00:25:47,080 --> 00:25:50,920 JUST ENGINEERED SOME OF THESE 643 00:25:50,920 --> 00:25:52,920 GUIDE RNAs ACCORDING TO 644 00:25:52,920 --> 00:25:53,440 ORIGINAL STRAIN. 645 00:25:53,440 --> 00:25:55,320 THIS IS A CLASSICAL REACTION 646 00:25:55,320 --> 00:25:58,240 WHERE YOU BASICALLY PUT ALL THE 647 00:25:58,240 --> 00:25:59,240 COMPONENTS TOGETHER AND YOU CAN 648 00:25:59,240 --> 00:26:03,600 SEE THAT IF YOU ADD THE SARS 649 00:26:03,600 --> 00:26:04,880 COV-2 VIRAL RNA YOU CAN SEE 650 00:26:04,880 --> 00:26:07,440 FLUORESCENCE IS TAKING OFF. 651 00:26:07,440 --> 00:26:09,320 IF YOU DON'T ADD IT YOU HAVE 652 00:26:09,320 --> 00:26:12,720 CERTAIN BACKGROUND OF JUST BY 653 00:26:12,720 --> 00:26:15,040 THE REPORTER AND THE ENZYME BY 654 00:26:15,040 --> 00:26:16,080 ITSELF. 655 00:26:16,080 --> 00:26:18,200 YOU SEE THAT THERE IS -- THE 656 00:26:18,200 --> 00:26:20,520 GUIDES THAT WORK VERY WELL, AND 657 00:26:20,520 --> 00:26:22,080 THERE ARE GUIDES THAT WORK NOT 658 00:26:22,080 --> 00:26:22,560 VERY WELL. 659 00:26:22,560 --> 00:26:25,120 THIS IS LITERALLY THE FIRST 15 660 00:26:25,120 --> 00:26:26,320 GUIDE RNAS WE DEVELOPED TWO 661 00:26:26,320 --> 00:26:28,880 YEARS AGO. 662 00:26:28,880 --> 00:26:30,200 WHAT WE FOUND IS IT WAS TRUE 663 00:26:30,200 --> 00:26:32,240 WHAT OTHERS FOUND BEFORE IF WE 664 00:26:32,240 --> 00:26:34,720 USE JUST ONE GUIDE AND THE 665 00:26:34,720 --> 00:26:36,440 ENZYME, THE SENSITIVITY IS NOT 666 00:26:36,440 --> 00:26:38,360 VERY HIGH AND WE CANNOT GET INTO 667 00:26:38,360 --> 00:26:41,120 A RANGE THAT IS ATTRACTIVE FOR 668 00:26:41,120 --> 00:26:42,160 ANY DIAGNOSTIC. 669 00:26:42,160 --> 00:26:44,280 HOWEVER WHEN WE START COMBINE 670 00:26:44,280 --> 00:26:47,880 GUIDES AND BUILD GUIDES ALONG 671 00:26:47,880 --> 00:26:50,320 THE VIRAL GENOME WE CAN VERY 672 00:26:50,320 --> 00:26:53,520 MUCH ENHANCE OUR SENSITIVITY 673 00:26:53,520 --> 00:26:56,200 HERE IN SHORTER AND SHORTER TIME 674 00:26:56,200 --> 00:26:56,680 FRAMES. 675 00:26:56,680 --> 00:26:59,680 I THINK ONE OF THE THINGS WE 676 00:26:59,680 --> 00:27:00,880 ALSO DID WHICH IS DIFFERENT FROM 677 00:27:00,880 --> 00:27:03,160 OTHERS IS WE DON'T LOOK AT END 678 00:27:03,160 --> 00:27:05,280 POINT, WE ARE NOT WAITING 30 679 00:27:05,280 --> 00:27:06,480 MINUTES OR TWO HOURS AND 680 00:27:06,480 --> 00:27:08,160 MEASURING THE END POINT, WE 681 00:27:08,160 --> 00:27:10,280 DEVELOPED ALGORITHMS WHERE WE 682 00:27:10,280 --> 00:27:12,000 DEVELOPED -- WE MEASURE THE 683 00:27:12,000 --> 00:27:13,440 SLOPE OF THE REACTION AND THAT 684 00:27:13,440 --> 00:27:16,920 LETS US VERY EARLY ON DETERMINE 685 00:27:16,920 --> 00:27:18,480 THIS IS A POSITIVE REACTION AND 686 00:27:18,480 --> 00:27:19,880 THIS IS BASICALLY A NEGATIVE 687 00:27:19,880 --> 00:27:23,000 REACTION HERE. 688 00:27:23,000 --> 00:27:27,040 SO WITH THIS WE HAVE BEEN 689 00:27:27,040 --> 00:27:28,360 COMBINING NOW MANY GUIDES, WE 690 00:27:28,360 --> 00:27:30,680 HAVE COME UP WITH AN EIGHT GUIDE 691 00:27:30,680 --> 00:27:35,480 COMBINATION IN OUR PAPER END OF 692 00:27:35,480 --> 00:27:38,200 2020, WE TALKED ABOUT FREE 693 00:27:38,200 --> 00:27:40,120 GUIDES BUT WE HAVE SINCE SETTLED 694 00:27:40,120 --> 00:27:41,680 ON AN EIGHT GUIDE COMBINATION 695 00:27:41,680 --> 00:27:44,120 THAT WE HAVE TESTED AGAINST ALL 696 00:27:44,120 --> 00:27:45,680 RISING VARIANTS AND THAT DETECTS 697 00:27:45,680 --> 00:27:49,600 IT INCLUDING OMICRON AND DELTA 698 00:27:49,600 --> 00:27:51,480 BUT NO CROSS REACTIVITY WITH 699 00:27:51,480 --> 00:27:55,520 NON-SARS COV-2 VIRAL RNA AND 700 00:27:55,520 --> 00:27:57,880 IMPORTANTLY ALSO NO CROSS 701 00:27:57,880 --> 00:28:01,280 REACTIVITY WITH HUMAN RNA AND 702 00:28:01,280 --> 00:28:04,560 ANY PREDICTABLE MICROBIOME RNA 703 00:28:04,560 --> 00:28:05,960 THAT WE MIGHT HAVE IN OUR NOSE. 704 00:28:05,960 --> 00:28:08,360 WITH THIS EIGHT GUIDE 705 00:28:08,360 --> 00:28:09,920 COMBINATION AND SOME OTHER 706 00:28:09,920 --> 00:28:12,440 TWEAKS THAT I WILL PARTIALLY 707 00:28:12,440 --> 00:28:13,840 ELUDE TO IN MY TALK, WE HAVE NOW 708 00:28:13,840 --> 00:28:16,480 PUSHED THE LIMIT OF DETECTION 709 00:28:16,480 --> 00:28:19,600 DOWN TO TEN COPIES PER 710 00:28:19,600 --> 00:28:21,640 MICROLITER, YOU CAN SEE HERE 711 00:28:21,640 --> 00:28:25,280 AFTER 20 MINUTES WE CAN CALL 712 00:28:25,280 --> 00:28:27,080 THIS REACTION POSITIVE WITHOUT 713 00:28:27,080 --> 00:28:29,360 ANY DOUBT. 714 00:28:29,360 --> 00:28:32,520 OVER THE GREEN LINE OF THE 715 00:28:32,520 --> 00:28:33,640 NEGATIVE AND ONE OF THE MOST 716 00:28:33,640 --> 00:28:34,440 IMPORTANT THINGS WE HAVE TO DO 717 00:28:34,440 --> 00:28:36,320 IS REALLY REDUCE THE BACKGROUND 718 00:28:36,320 --> 00:28:39,080 HERE IN ORDER TO HAVE THE 719 00:28:39,080 --> 00:28:41,440 SENSITIVITY TO DIFFERENTIATE 720 00:28:41,440 --> 00:28:44,280 BETWEEN THESE SMALL DIFFERENCES 721 00:28:44,280 --> 00:28:45,280 IN SIGNAL. 722 00:28:45,280 --> 00:28:47,000 ANOTHER THING THAT TURNED OUT TO 723 00:28:47,000 --> 00:28:49,480 BE VERY IMPORTANT FOR THE 724 00:28:49,480 --> 00:28:52,720 SUCCESS OF THIS ESSAY WAS THE 725 00:28:52,720 --> 00:28:55,200 OPTICS. 726 00:28:55,200 --> 00:28:56,720 INITIALLY WE THOUGHT WHILE IS 727 00:28:56,720 --> 00:28:58,480 GOAL IS MOBILE PHONE WILL BE AS 728 00:28:58,480 --> 00:29:01,360 SENSITIVE AS THE PLATE READER IN 729 00:29:01,360 --> 00:29:03,000 THE LAB, IT TURNS OUT THAT THE 730 00:29:03,000 --> 00:29:04,880 MOBILE PHONE CAMERAS THAT WE 731 00:29:04,880 --> 00:29:07,640 HAVE MOST OF OUR PHONES TODAY 732 00:29:07,640 --> 00:29:09,440 ARE ACTUALLY TEN TIMES BETTER 733 00:29:09,440 --> 00:29:10,480 THAN THE PLATE READER WE HAVE IN 734 00:29:10,480 --> 00:29:10,920 THE LAB. 735 00:29:10,920 --> 00:29:14,080 THIS IS MAINLY BECAUSE OF THE 736 00:29:14,080 --> 00:29:15,680 NOISINESS OF THE SIGNAL, THE 737 00:29:15,680 --> 00:29:19,080 PLATE READER HAS A CONSIDERABLE 738 00:29:19,080 --> 00:29:24,480 NOISE IN READS WHILE HERE THE 739 00:29:24,480 --> 00:29:26,680 PIXEL 4 PHONE CAMERA IS STEADY 740 00:29:26,680 --> 00:29:29,000 IN ACQUIRING THE SIGNALS AND 741 00:29:29,000 --> 00:29:31,280 GIVES US OVERALL TENFOLD 742 00:29:31,280 --> 00:29:34,680 INCREASE IN IF SENSITIVITY WHICH 743 00:29:34,680 --> 00:29:35,720 IS REMARKABLE. 744 00:29:35,720 --> 00:29:39,360 THIS IS NOT DUE ONLY TO 745 00:29:39,360 --> 00:29:40,400 EXPENSIVE PHONES THAT WE HAVE 746 00:29:40,400 --> 00:29:41,880 CARRYING IN OUR POCKETS. 747 00:29:41,880 --> 00:29:44,680 WE HAVE TESTED $20 PHONES FROM 748 00:29:44,680 --> 00:29:50,880 INDIA AND THEY ALSO HAVE PHONE 749 00:29:50,880 --> 00:29:51,800 CAMERAS IN THE RANGE OF 750 00:29:51,800 --> 00:29:52,680 SENSITIVITY THAT IS IMPORTANT 751 00:29:52,680 --> 00:29:55,040 FOR US. 752 00:29:55,040 --> 00:29:57,840 SO FIRST MODEL WE BUILT THAT DAN 753 00:29:57,840 --> 00:30:00,840 BUILT WAS BASICALLY THE PHONE 754 00:30:00,840 --> 00:30:03,800 HERE, ALIGN, THE CAMERA ALIGNED 755 00:30:03,800 --> 00:30:08,680 TO THIS BOX WHICH CARRIES THE 756 00:30:08,680 --> 00:30:09,200 REACTION. 757 00:30:09,200 --> 00:30:11,040 THERE IS A DRAWER HERE YOU CAN 758 00:30:11,040 --> 00:30:14,320 OPEN AND ENTER THE REACTION AND 759 00:30:14,320 --> 00:30:16,800 BASICALLY THIS IS A PROGRAM, AN 760 00:30:16,800 --> 00:30:18,240 APP THAT HAS BEEN WRITTEN FOR 761 00:30:18,240 --> 00:30:20,280 THE PHONE THAT IMMEDIATELY 762 00:30:20,280 --> 00:30:22,560 ACQUIRES FLUORESCENCE THAT IS IN 763 00:30:22,560 --> 00:30:23,680 THESE THREE CHANNELS HERE AND 764 00:30:23,680 --> 00:30:25,480 YOU CAN SEE THIS IS A VERY 765 00:30:25,480 --> 00:30:28,280 HIGHLY POSITIVE PATIENT SAMPLE, 766 00:30:28,280 --> 00:30:30,200 LESS POSITIVE PATIENT SAMPLE IN 767 00:30:30,200 --> 00:30:32,640 NEGATIVE ONE AND YOU CAN SEE 768 00:30:32,640 --> 00:30:35,840 THAT THE PHONE IS BASICALLY 769 00:30:35,840 --> 00:30:37,920 RUNNING THE REACTION, THE 770 00:30:37,920 --> 00:30:39,160 ACQUISITION AND ANALYSIS AND I 771 00:30:39,160 --> 00:30:43,120 THINK THE PHONE HAS THE ADDED 772 00:30:43,120 --> 00:30:47,080 BENEFIT IT ALSO IS CONNECTED TO 773 00:30:47,080 --> 00:30:50,080 LOCATION, GPS SYSTEMS SO IT CAN 774 00:30:50,080 --> 00:30:54,280 BE USED TO POTENTIALLY IN THE 775 00:30:54,280 --> 00:30:56,280 FUTURE FOR GEOLOCATION AND O 776 00:30:56,280 --> 00:30:59,840 CONTACT TRACING IF WE GO FORWARD 777 00:30:59,840 --> 00:31:02,320 WITH THIS TECHNOLOGY. 778 00:31:02,320 --> 00:31:05,760 NOW, THIS -- ADDING MULTIPLE 779 00:31:05,760 --> 00:31:07,440 GUIDES, IMPROVING THE OPTICS, 780 00:31:07,440 --> 00:31:11,840 HAS REALLY LED TO EXQUISITE 781 00:31:11,840 --> 00:31:14,840 SENSITIVITY OF THAT'S SAY WE ARE 782 00:31:14,840 --> 00:31:16,440 IN OUR INITIAL PAPER WE COULD 783 00:31:16,440 --> 00:31:17,680 SHOW WE HAVE HAD FIVE PATIENT 784 00:31:17,680 --> 00:31:21,760 SAMPLES FROM THE BIOHUB HERE IN 785 00:31:21,760 --> 00:31:24,480 SAN FRANCISCO AND WE CAN DETECT 786 00:31:24,480 --> 00:31:26,040 ALL FIVE PATIENT SAMPLES WITHIN 787 00:31:26,040 --> 00:31:29,480 FIVE MINUTES AS POSITIVE AND 788 00:31:29,480 --> 00:31:30,680 DISTINGUISH IT FROM THE 789 00:31:30,680 --> 00:31:31,440 NEGATIVES. 790 00:31:31,440 --> 00:31:33,720 SO THIS IS VERY PROMISING. 791 00:31:33,720 --> 00:31:35,720 AND WE CONTINUING TO WORK ON 792 00:31:35,720 --> 00:31:38,160 THIS SO-CALLED BULK ASSAY AS WE 793 00:31:38,160 --> 00:31:40,760 USE IT WITH THE EIGHT GUIDES 794 00:31:40,760 --> 00:31:42,240 THAT I HAVE INTRODUCED TO YOU. 795 00:31:42,240 --> 00:31:45,320 WE ARE LOOKING BUILDING DEVICES 796 00:31:45,320 --> 00:31:48,080 LIKE THIS, WHERE WE PUT THE SWAB 797 00:31:48,080 --> 00:31:50,440 INTO A PEN LIKE STRUCTURE, SCREW 798 00:31:50,440 --> 00:31:54,400 IT INTO THIS DETECTION BLOCK, 799 00:31:54,400 --> 00:31:56,640 THE IMPORTANT PART IS THAT WE 800 00:31:56,640 --> 00:31:59,080 HAVE TO HEAT, STILL HEAT AND 801 00:31:59,080 --> 00:32:00,640 THAT IS ONE THING WE WOULD LIKE 802 00:32:00,640 --> 00:32:02,720 TO GET RID OF BUT WE HAVE TO 803 00:32:02,720 --> 00:32:04,280 BRIEFLY HEAT THE REACTION, 804 00:32:04,280 --> 00:32:07,080 MAINLY TO INACTIVATE RNA, 805 00:32:07,080 --> 00:32:09,080 BECAUSE THAT IS OUR MAIN ENANY 806 00:32:09,080 --> 00:32:12,920 IN THIS ESSAY AND THEN WE CAN 807 00:32:12,920 --> 00:32:14,200 TURN AND PUSH REACTIONS TO THE 808 00:32:14,200 --> 00:32:16,120 THREE CHANNELS, IT IS A POSITIVE 809 00:32:16,120 --> 00:32:20,840 AND NEGATIVE AND THE TESTING 810 00:32:20,840 --> 00:32:21,200 CHANNEL. 811 00:32:21,200 --> 00:32:23,080 THEN YOU CAN USE YOUR MOBILE 812 00:32:23,080 --> 00:32:25,520 PHONE BASICALLY ALIGNED WITH A 813 00:32:25,520 --> 00:32:30,880 CAMERA HERE, TO READ THIS WITH 814 00:32:30,880 --> 00:32:31,480 APPROPRIATE APP. 815 00:32:31,480 --> 00:32:37,880 SO THAT IS THE SORT OF THE BROAD 816 00:32:37,880 --> 00:32:39,080 STROKE ROUTE WE ARE CURRENTLY 817 00:32:39,080 --> 00:32:39,600 GOING. 818 00:32:39,600 --> 00:32:41,520 WE HAVE BEEN VERY FRUSTRATED, WE 819 00:32:41,520 --> 00:32:42,600 THOUGHT WE COULD DO THIS MUCH 820 00:32:42,600 --> 00:32:44,880 FASTER AND COULD BE READY FOR 821 00:32:44,880 --> 00:32:46,520 SOMETHING LIKE THE OMICRON SURGE 822 00:32:46,520 --> 00:32:47,760 HERE BUT BUILDING SOMETHING FROM 823 00:32:47,760 --> 00:32:50,760 SCRATCH AND HAVING FDA APPROVAL 824 00:32:50,760 --> 00:32:52,760 IS NOT EASY BUT WE THINK THAT 825 00:32:52,760 --> 00:32:55,520 THIS IS A TECHNOLOGY THAT IS NOT 826 00:32:55,520 --> 00:32:57,080 ONLY IMPORTANT NOW FOR SARS 827 00:32:57,080 --> 00:32:59,000 COV-2 BUT ALSO FOR OTHER 828 00:32:59,000 --> 00:33:00,200 VIRUSES, RNA VIRUSES IN THE 829 00:33:00,200 --> 00:33:02,400 FUTURE AND ALSO I THINK IS A 830 00:33:02,400 --> 00:33:03,680 METHOD THAT IS EXTREMELY 831 00:33:03,680 --> 00:33:05,400 INTERESTING FOR RNA 832 00:33:05,400 --> 00:33:08,200 QUANTIFICATION ALSO IN THE LAB. 833 00:33:08,200 --> 00:33:11,680 SO THIS WORK WAS MAINLY DRIVEN 834 00:33:11,680 --> 00:33:15,200 BY VERY TALENTED MSTP STUDENT IN 835 00:33:15,200 --> 00:33:19,920 THE LAB PARINAZ FOZOUNI, SHE IS 836 00:33:19,920 --> 00:33:22,320 BACK IN MEDICAL SCHOOL, SHE 837 00:33:22,320 --> 00:33:26,000 WORKED WITH SUNGMIN SON WHO HAS 838 00:33:26,000 --> 00:33:28,400 HELPED WITH OPTICS AND THE 839 00:33:28,400 --> 00:33:30,920 ACQUISITION OF -- WITH THE 840 00:33:30,920 --> 00:33:33,160 MOBILE PHONE. 841 00:33:33,160 --> 00:33:35,080 SUNGMIN HAS TAKEN THE NEXT STEP 842 00:33:35,080 --> 00:33:37,080 AND PUT IT INTO A DROPLET BASED 843 00:33:37,080 --> 00:33:37,440 WORK FLOW. 844 00:33:37,440 --> 00:33:39,040 BECAUSE OF THE -- I TOLD YOU WE 845 00:33:39,040 --> 00:33:41,960 HAVE WITH THE BULK ESSAY 846 00:33:41,960 --> 00:33:43,760 ACHIEVED A UNIQUE SENSITIVITY 847 00:33:43,760 --> 00:33:46,200 BUT WE REALLY WOULD LIKE TO GO 848 00:33:46,200 --> 00:33:48,480 INTO PCR SENSITIVITY WHICH IS 849 00:33:48,480 --> 00:33:51,800 ONE COPY FROM MICROLITER BUT 850 00:33:51,800 --> 00:33:53,680 ALSO LIKE TO DECREASE NUMBER OF 851 00:33:53,680 --> 00:33:57,360 GUIDES IN ORDER TO BE ABLE TO 852 00:33:57,360 --> 00:34:00,080 DIFFERENTIATE BETWEEN INDIVIDUAL 853 00:34:00,080 --> 00:34:02,320 VARIANTS OR VIRUSES. 854 00:34:02,320 --> 00:34:05,080 SO WHAT HE DID IS VERY SIMPLE, 855 00:34:05,080 --> 00:34:08,480 HE EMULSIFIED THE REACTION BY 856 00:34:08,480 --> 00:34:10,200 PIPETTING UP AND DOWN AND 857 00:34:10,200 --> 00:34:11,400 BUILDING THESE DROPLETS AND 858 00:34:11,400 --> 00:34:12,720 IMAGING AND QUANTIFYING THE 859 00:34:12,720 --> 00:34:16,360 FLUORESCENCE AND THE INDIVIDUAL 860 00:34:16,360 --> 00:34:18,640 DROPLETS BY MICROSCOPY. 861 00:34:18,640 --> 00:34:19,880 YOU CAN SEE WE HAVE DIFFERENT 862 00:34:19,880 --> 00:34:21,960 SIZES OF DROPLETS BUT BECAUSE WE 863 00:34:21,960 --> 00:34:24,400 ARE USING MICROSCOPY NORMALIZED 864 00:34:24,400 --> 00:34:25,960 FOR THIS, YOU CAN SEE WITHIN A 865 00:34:25,960 --> 00:34:28,880 FEW MINUTES, YOU SEE THE 866 00:34:28,880 --> 00:34:30,440 DROPLETS POPPING UP AND NUMBER 867 00:34:30,440 --> 00:34:34,280 OF DROPLETS IS BASICALLY THE 868 00:34:34,280 --> 00:34:36,280 QUANTITATION OF THIS ASSAY. 869 00:34:36,280 --> 00:34:37,960 WHILE YOU CAN SEE OVER 30 870 00:34:37,960 --> 00:34:39,280 MINUTES WE HAVE AN INCREASE, 871 00:34:39,280 --> 00:34:42,280 CONTINUED INCREASE IN THE SIGNAL 872 00:34:42,280 --> 00:34:44,360 PER DROP, THE NUMBER OF DROPS 873 00:34:44,360 --> 00:34:45,800 REMAINS STEADY AFTER FIVE 874 00:34:45,800 --> 00:34:48,800 MINUTES SO WE CAN BASICALLY CALL 875 00:34:48,800 --> 00:34:51,120 THIS REACTION AFTER FIVE MINUTES 876 00:34:51,120 --> 00:34:53,320 WHICH IS A HUGE BENEFIT. 877 00:34:53,320 --> 00:34:55,680 AND WE HAVE A HUGE BENEFIT IN 878 00:34:55,680 --> 00:34:57,240 SENSITIVITY BECAUSE WE NOW PULL 879 00:34:57,240 --> 00:35:00,200 FROM A 300-MICROLITER REACTION 880 00:35:00,200 --> 00:35:02,480 TO A 30 PICO LITTER REACTION IN 881 00:35:02,480 --> 00:35:02,960 A DROPLET. 882 00:35:02,960 --> 00:35:06,120 YOU CAN SEE THAT WITH ONE GUIDE 883 00:35:06,120 --> 00:35:11,000 WE CAN NOW DETECT 20 OR 10 COP 884 00:35:11,000 --> 00:35:13,680 PIPS PER MICROLITER WHICH IS THE 885 00:35:13,680 --> 00:35:14,600 SENSITIVITY WITH EIGHT GUIDES IN 886 00:35:14,600 --> 00:35:16,800 THE BULK REACTIONND AND IF WE 887 00:35:16,800 --> 00:35:18,160 COMBINE THE GUIDES HERE IN 888 00:35:18,160 --> 00:35:19,400 DIFFERENT VARIATIONS WE CAN GO 889 00:35:19,400 --> 00:35:22,120 TO ONE COPY PER MICROLITER OR 890 00:35:22,120 --> 00:35:23,800 BELOW, EASILY IN FIVE MINUTES. 891 00:35:23,800 --> 00:35:27,080 SO THIS IS REALLY A UNIQUELY 892 00:35:27,080 --> 00:35:28,880 SENSITIVE METHOD THAT PUSHES 893 00:35:28,880 --> 00:35:30,400 THIS DIRECT DETECTION METHOD 894 00:35:30,400 --> 00:35:34,400 INTO A PCR SENSITIVITY. 895 00:35:34,400 --> 00:35:37,080 BUT THE INTERESTING PART, OR THE 896 00:35:37,080 --> 00:35:41,520 PART WITH THE MOST INTERESTING 897 00:35:41,520 --> 00:35:44,560 IS THAT THE DROPLETS OFFER US 898 00:35:44,560 --> 00:35:50,160 NOW A WAY TO DIFFERENTIATE 899 00:35:50,160 --> 00:35:51,440 BETWEEN INDIVIDUAL GUIDES AND 900 00:35:51,440 --> 00:35:53,960 THERE BY GIVES US A WAY TO 901 00:35:53,960 --> 00:35:55,280 POTENTIALLY MEASURE DIFFERENT 902 00:35:55,280 --> 00:35:56,720 VIRUSES OR DIFFERENT VARIANTS 903 00:35:56,720 --> 00:35:58,040 JUST IN ONE SINGLE REACTION A. 904 00:35:58,040 --> 00:36:00,040 INSTEAD OF PUTTING THEM INTO THE 905 00:36:00,040 --> 00:36:01,400 THREE CHANNELS OR IN MORE 906 00:36:01,400 --> 00:36:04,320 CHANNELS AND DIVIDING OUR INPUT 907 00:36:04,320 --> 00:36:06,840 MATERIAL, I THINK WE CAN NOW USE 908 00:36:06,840 --> 00:36:08,920 ONE SINGLE REACTION AND CAN 909 00:36:08,920 --> 00:36:10,640 POTENTIALLY COMBINE MULTIPLE 910 00:36:10,640 --> 00:36:11,120 GUIDES. 911 00:36:11,120 --> 00:36:14,040 HOW -- WHY CAN WE DO THAT? 912 00:36:14,040 --> 00:36:15,280 THE REASON IS SIMPLE. 913 00:36:15,280 --> 00:36:17,240 IF YOU REMEMBER MY VERY FIRST 914 00:36:17,240 --> 00:36:18,560 SLIDE I SHOWED YOU ALL THE 915 00:36:18,560 --> 00:36:23,040 DIFFERENT GUIDE KINETICS, EACH 916 00:36:23,040 --> 00:36:25,120 OF THEM HAS -- WE CALL THEM THE 917 00:36:25,120 --> 00:36:28,080 GOOD GUIDES OR THE BAD GUIDES. 918 00:36:28,080 --> 00:36:31,400 VERY STEEP SLOPE WHERE THEY HAVE 919 00:36:31,400 --> 00:36:36,240 A VERY SORT OF SHALLOW SLOPE. 920 00:36:36,240 --> 00:36:38,960 BUT THIS SLOPE CHARACTERISTIC IN 921 00:36:38,960 --> 00:36:41,480 THE DROPLETS IS FIXED BECAUSE WE 922 00:36:41,480 --> 00:36:46,480 ALWAYS HAVE ONE COPY OF THE 923 00:36:46,480 --> 00:36:47,560 TARGET. 924 00:36:47,560 --> 00:36:52,840 SO WE CAN USE THIS KINETIC SLOPE 925 00:36:52,840 --> 00:36:55,280 BEHAVIOR OF THE INDIVIDUAL GUIDE 926 00:36:55,280 --> 00:36:56,960 TO BAR CODE AND DIFFERENTIATE 927 00:36:56,960 --> 00:36:58,800 BETWEEN INDIVIDUAL GUIDES. 928 00:36:58,800 --> 00:37:01,040 I AM SHOWING AN EXAMPLE HERE, IF 929 00:37:01,040 --> 00:37:03,440 WE HAVE A GUIDE THAT IS SPECIFIC 930 00:37:03,440 --> 00:37:04,800 FOR VARIANT, THIS IS THE 931 00:37:04,800 --> 00:37:07,000 CALIFORNIA VARIANT HERE WITH THE 932 00:37:07,000 --> 00:37:10,600 SIGNAL POINT MUTATION. 933 00:37:10,600 --> 00:37:13,600 THIS IF THAT GUIDE YIELDS TO THE 934 00:37:13,600 --> 00:37:16,640 VARIANT RNA IT HAS A STEEPER 935 00:37:16,640 --> 00:37:18,640 SLOPE THAN TO THE WILD TYPE 936 00:37:18,640 --> 00:37:21,080 WHERE IT IS INCOMPLETE. 937 00:37:21,080 --> 00:37:22,880 THIS DIFFERENCE IN SLOPE CAN NOW 938 00:37:22,880 --> 00:37:26,080 BE USED BY ALLIES IN PATIENT 939 00:37:26,080 --> 00:37:27,680 SAMPLES WHERE WE CAN 940 00:37:27,680 --> 00:37:29,880 DIFFERENTIATE WHAT THIS ONE 941 00:37:29,880 --> 00:37:33,520 GUIDE WITH ALMOST 100% ACCURACY 942 00:37:33,520 --> 00:37:34,600 BETWEEN WILD TYPE SAMPLES THAT 943 00:37:34,600 --> 00:37:38,160 WE HAVE IN PATIENTS, OR BETWEEN 944 00:37:38,160 --> 00:37:39,360 CALIFORNIA VARIANT THAT WAS 945 00:37:39,360 --> 00:37:41,480 WIDESPREAD HERE IN SAN 946 00:37:41,480 --> 00:37:41,880 FRANCISCO. 947 00:37:41,880 --> 00:37:43,800 IN OUR SAMPLES. 948 00:37:43,800 --> 00:37:46,360 WE ARE VERY EXCITED BY THIS 949 00:37:46,360 --> 00:37:47,800 KINETIC BAR CODING HERE BECAUSE 950 00:37:47,800 --> 00:37:51,680 THAT ALLOWS US NOW TO MULTIPLEX 951 00:37:51,680 --> 00:37:53,680 REACTION, TRULY MULTIPLEX THE 952 00:37:53,680 --> 00:37:54,960 REACTION IN ONE REACTION WHERE 953 00:37:54,960 --> 00:37:57,520 WE CAN NOW BASED ON KINETICS ON 954 00:37:57,520 --> 00:38:00,960 THE INDIVIDUAL OF THE INDIVIDUAL 955 00:38:00,960 --> 00:38:02,040 GUIDES DIFFERENTIATE BETWEEN 956 00:38:02,040 --> 00:38:04,040 VARIANTS BUT ALSO POTENTIALLY 957 00:38:04,040 --> 00:38:08,000 LOOK FOR MULTIPLE VIRUSES IN ONE 958 00:38:08,000 --> 00:38:08,920 REACTION. 959 00:38:08,920 --> 00:38:12,760 SO AS A SUMMARY I HAVE SHOWN YOU 960 00:38:12,760 --> 00:38:14,320 DIRECT DETECTION WITH CRISPR 961 00:38:14,320 --> 00:38:17,680 WITH CAS 13A IN BULK OR DROPLET 962 00:38:17,680 --> 00:38:19,160 REACTIONS IS POSSIBLE AND I 963 00:38:19,160 --> 00:38:21,360 EMPHASIZE HERE THIS IS THE 964 00:38:21,360 --> 00:38:23,480 DIRECT ROUTE TO DETECTION ESSAY, 965 00:38:23,480 --> 00:38:25,680 NOT AMPLIFIED REACTION AS IN 966 00:38:25,680 --> 00:38:27,680 OTHER CRISPR ASSAYS. 967 00:38:27,680 --> 00:38:30,200 THE SENSITIVITY IS CURRENTLY TEN 968 00:38:30,200 --> 00:38:31,880 COPIES PER MICROLITER AND 30 969 00:38:31,880 --> 00:38:33,800 MINUTES BULK REACTION OR ONE 970 00:38:33,800 --> 00:38:35,280 COPY PER MICROLITER IN FIVE 971 00:38:35,280 --> 00:38:36,040 MINUTES FOR THE DROPLET 972 00:38:36,040 --> 00:38:38,920 REACTION. 973 00:38:38,920 --> 00:38:41,040 THE DROPLETS OFFER UNIQUE 974 00:38:41,040 --> 00:38:41,880 OPPORTUNITIES BECAUSE NOW WE 975 00:38:41,880 --> 00:38:43,240 HAVE THE SINGLE MOLECULE 976 00:38:43,240 --> 00:38:45,120 RESOLUTION IN THE DROPLETS THAT 977 00:38:45,120 --> 00:38:48,880 ALLOWS US TO STUDY THE BASIC 978 00:38:48,880 --> 00:38:51,120 BIOLOGY OF CAS 13A BUT ALSO 979 00:38:51,120 --> 00:38:53,360 ALLOWS KINETIC BAR CODING BY 980 00:38:53,360 --> 00:38:55,160 USING THE UNIQUE INTERACTION OR 981 00:38:55,160 --> 00:38:59,600 THE UNIQUE KINETICS THAT RESULTS 982 00:38:59,600 --> 00:39:00,280 FROM THE GUIDE TARGET 983 00:39:00,280 --> 00:39:01,840 INTERACTION. 984 00:39:01,840 --> 00:39:04,480 THIS ALLOWS US TO MULTIPLEX 985 00:39:04,480 --> 00:39:05,760 DIFFERENT TARGETS WITH SINGLE 986 00:39:05,760 --> 00:39:06,560 REPORTSER WE DON'T HAVE TO 987 00:39:06,560 --> 00:39:07,880 CHANGE THE ENZYME, THE REPORTER 988 00:39:07,880 --> 00:39:08,960 OR ANYTHING. 989 00:39:08,960 --> 00:39:11,680 AND IT GIVES UNIQUE, THAT I 990 00:39:11,680 --> 00:39:13,280 HAVEN'T EMPHASIZED BUT THIS IS A 991 00:39:13,280 --> 00:39:15,960 UNIQUE QUANTITATIVE ASSAY 992 00:39:15,960 --> 00:39:17,880 BECAUSE OUR SLOPE IS DIRECTLY 993 00:39:17,880 --> 00:39:19,320 LINEARLY RELATED TO THE VIRAL 994 00:39:19,320 --> 00:39:21,120 LOAD IN OUR ASSAY. 995 00:39:21,120 --> 00:39:25,400 SO WE CAN DIRECTLY FROM THE 996 00:39:25,400 --> 00:39:28,240 SLOPE DERIVE QUANTITATION OF 997 00:39:28,240 --> 00:39:31,640 VIRAL LOADS IN OUR ASSAY SO WE 998 00:39:31,640 --> 00:39:33,320 CAN HAVE ACCURATE FAST MOBILE 999 00:39:33,320 --> 00:39:34,800 RNA DETECTION THAT WE THINK IS 1000 00:39:34,800 --> 00:39:37,800 IMPORTANT FOR SARS COV-2 IN THIS 1001 00:39:37,800 --> 00:39:39,600 PANDEMIC BUT POTENTIALLY OTHER 1002 00:39:39,600 --> 00:39:41,640 RNAS NOT ONLY VIRAL RNAs IN 1003 00:39:41,640 --> 00:39:44,280 THE FUTURE. 1004 00:39:44,280 --> 00:39:47,520 THIS LEADS ME TO MY LAST PART OF 1005 00:39:47,520 --> 00:39:48,440 THE TALK WHICH IS THE STUDY 1006 00:39:48,440 --> 00:39:50,680 PART. 1007 00:39:50,680 --> 00:39:52,680 AND THIS IS CONTINUED 1008 00:39:52,680 --> 00:39:54,920 COLLABORATION BETWEEN JENNIFER'S 1009 00:39:54,920 --> 00:39:56,840 LAB AND MY LAB. 1010 00:39:56,840 --> 00:39:59,160 JENNIFER MOVED 25% OF HER 1011 00:39:59,160 --> 00:40:01,160 EFFORTS A FEW YEARS BACK TO 1012 00:40:01,160 --> 00:40:04,760 GLADSTONE MAINLY ALSO TO HAVE AT 1013 00:40:04,760 --> 00:40:09,880 THE UCSF CAMPUS AND HAVE A MORE 1014 00:40:09,880 --> 00:40:17,000 CLINICALLY AFFILIATED LAB HERE 1015 00:40:17,000 --> 00:40:18,280 IN SAN FRANCISCO. 1016 00:40:18,280 --> 00:40:20,760 ONE OF HER POST-DOCS IS 1017 00:40:20,760 --> 00:40:21,760 EXTREMELY TALENTED AND HER LAB 1018 00:40:21,760 --> 00:40:23,360 IS NEXT TO MINE SO WE HAD 1019 00:40:23,360 --> 00:40:25,600 ENORMOUS INTERACTIONS DURING OUR 1020 00:40:25,600 --> 00:40:29,040 TIME FOR THE DEVICE AND THE TEST 1021 00:40:29,040 --> 00:40:30,480 BUT WE HAVE ALSO PEOPLE IN HER 1022 00:40:30,480 --> 00:40:32,880 LAB AND MY LAB HAVE CONTINUED TO 1023 00:40:32,880 --> 00:40:34,840 WORK CLOSELY TOGETHER AND 1024 00:40:34,840 --> 00:40:38,240 AMAZING SYNERGIES HAVE BEEN 1025 00:40:38,240 --> 00:40:39,160 ARISING. 1026 00:40:39,160 --> 00:40:41,480 SO SYED IS WORKING CLOSE WITH 1027 00:40:41,480 --> 00:40:44,800 VERY TALENTED POST DOC TAHA AND 1028 00:40:44,800 --> 00:40:47,440 MY LAB AND TAKAKO, A SENIOR 1029 00:40:47,440 --> 00:40:49,600 SCIENTIST WHO IS DOING ALL THE 1030 00:40:49,600 --> 00:40:53,040 BSL 3 WORK FOR THEM. 1031 00:40:53,040 --> 00:40:55,080 THIS LED IN NOVEMBER LAST YEAR 1032 00:40:55,080 --> 00:40:58,680 TO THE SCIENCE PAPER WHERE WE 1033 00:40:58,680 --> 00:41:01,920 DEVELOP NEW VIRUS LIKE PARTICLES 1034 00:41:01,920 --> 00:41:04,480 TO ASSESS SARS COV-2 IN BOTH 1035 00:41:04,480 --> 00:41:06,240 VARIANTS AND WE CONTINUED TO DO 1036 00:41:06,240 --> 00:41:09,280 THIS WITH DELTA AND NOW WITH 1037 00:41:09,280 --> 00:41:09,600 OMICRON. 1038 00:41:09,600 --> 00:41:12,160 WHAT IS A VIRUS LIKE PARTICLE 1039 00:41:12,160 --> 00:41:14,800 AND HOW IS IT DIFFERENT FROM A 1040 00:41:14,800 --> 00:41:17,920 PSEUDOTYPE VIRUS? 1041 00:41:17,920 --> 00:41:21,040 I THINK A VIRUS LIKE PARTICLE 1042 00:41:21,040 --> 00:41:24,280 USES ALL FOUR STRUCTURAL 1043 00:41:24,280 --> 00:41:27,880 PROTEINS OF SARS COV-2, NOT JUST 1044 00:41:27,880 --> 00:41:30,840 THE SPIKE PROTEIN THAT IS 1045 00:41:30,840 --> 00:41:33,360 BASICALLY ARTIFICIALLY MOUNTED 1046 00:41:33,360 --> 00:41:36,480 ON TO ANOTHER VIRUS PARTICLE. 1047 00:41:36,480 --> 00:41:40,480 WE ARE GENERATING AUTHENTIC SARS 1048 00:41:40,480 --> 00:41:41,600 COV-2 VIRUS PARTICLES INCLUDING 1049 00:41:41,600 --> 00:41:43,240 THE S PROTEIN, THE N PROTEIN, 1050 00:41:43,240 --> 00:41:46,320 THE NUCLEAR CAPSID, WHICH IS 1051 00:41:46,320 --> 00:41:48,680 CRITICAL FOR THE PARTICLE 1052 00:41:48,680 --> 00:41:51,320 FORMATION AND RNA INCORPORATION 1053 00:41:51,320 --> 00:41:53,640 BUT ALSO THE SMALLER ADDITIONAL 1054 00:41:53,640 --> 00:41:56,240 STRUCTURAL PROTEINS LIKE THE M 1055 00:41:56,240 --> 00:42:00,560 PROTEIN AND TINY LITTLE E 1056 00:42:00,560 --> 00:42:04,080 PROTEIN THAT CONSTITUENTS SARS 1057 00:42:04,080 --> 00:42:04,680 COV-2 PARTICLE. 1058 00:42:04,680 --> 00:42:08,440 IN ORDER TO USE A VIRAL LIKE 1059 00:42:08,440 --> 00:42:11,920 PARTICLE INSTEAD OF PSEUDOVIRUS, 1060 00:42:11,920 --> 00:42:14,360 SYED NEEDED TO IDENTIFY THE 1061 00:42:14,360 --> 00:42:16,360 PACKAGING SIGNAL FOR THIS 1062 00:42:16,360 --> 00:42:18,760 PARTICLE IN ORDER TO INCORPORATE 1063 00:42:18,760 --> 00:42:23,160 A REPORTER THAT WE COULD USE TO 1064 00:42:23,160 --> 00:42:24,320 MEASURE PRODUCTION AND 1065 00:42:24,320 --> 00:42:26,800 INFECTIVITY OF THIS PARTICLES. 1066 00:42:26,800 --> 00:42:29,760 HE DID THIS BY USING EXISTING 1067 00:42:29,760 --> 00:42:31,560 LITERATURE ABOUT CORONA VIRUS 1068 00:42:31,560 --> 00:42:36,560 PACKAGING SIGNALS, AND EXTENSIVE 1069 00:42:36,560 --> 00:42:38,280 MUTAGENESIS AND IDENTIFIED A 1070 00:42:38,280 --> 00:42:43,560 SMALL T 20 MOTIF AT THE END OF 1 1071 00:42:43,560 --> 00:42:45,880 AB AS PACKAGING SIGNAL AND IF HE 1072 00:42:45,880 --> 00:42:50,480 HOOKS UP TO GFP OR LUCIFERASE 1073 00:42:50,480 --> 00:42:54,680 GENE HE CAN SHOW THAT THIS 1074 00:42:54,680 --> 00:42:56,680 CONSTRUCT IS PACKAGED AND ALSO 1075 00:42:56,680 --> 00:42:58,960 DELIVERED INTO THE NEXT CELLS 1076 00:42:58,960 --> 00:43:01,720 WHERE EITHER GLOWS OR PRODUCES 1077 00:43:01,720 --> 00:43:04,040 LUCIFERASE. 1078 00:43:04,040 --> 00:43:06,360 SO WITH THIS AT HAND HE 1079 00:43:06,360 --> 00:43:10,320 DEVELOPED A VLP SYSTEM, SIMPLE 1080 00:43:10,320 --> 00:43:11,320 CO-TRANSFECTION SYSTEM WHERE YOU 1081 00:43:11,320 --> 00:43:12,680 USE THE SPIKE PROTEIN, THE N 1082 00:43:12,680 --> 00:43:17,040 PROTEIN EXPRESSING CONSTRUCT BUT 1083 00:43:17,040 --> 00:43:23,520 ALSO CISTRONIC AND EXPRESSING 1084 00:43:23,520 --> 00:43:28,120 REPORTER TO CO-TRANSFECT TO 2293 1085 00:43:28,120 --> 00:43:33,440 CELLS, YOU ISOLATE THEM PUT ON E 1086 00:43:33,440 --> 00:43:34,640 RECEPTOR CELLS AND THEN YOU CAN 1087 00:43:34,640 --> 00:43:37,280 MEASURE LUCIFERASE OR IF YOU USE 1088 00:43:37,280 --> 00:43:39,080 GFP YOU CAN DO SINGLE CELL 1089 00:43:39,080 --> 00:43:39,800 ANALYSIS BY FLOW. 1090 00:43:39,800 --> 00:43:42,520 THE IMPORTANT PART IS THAT THESE 1091 00:43:42,520 --> 00:43:44,680 VLPs ONLY FUNCTION IF YOU PUT 1092 00:43:44,680 --> 00:43:45,880 ALL FOUR TOGETHER IN THE RIGHT 1093 00:43:45,880 --> 00:43:46,720 RATIO. 1094 00:43:46,720 --> 00:43:49,440 IF YOU MISS OUT ON ANY OF THIS 1095 00:43:49,440 --> 00:43:51,560 YOU HAVE NO SIGNAL CONFIRMING 1096 00:43:51,560 --> 00:43:54,680 THAT YOU HAVE REALLY INTACT VLP 1097 00:43:54,680 --> 00:43:56,920 HERE IN SUPERRER NATANT. 1098 00:43:56,920 --> 00:43:58,280 WITH THIS WE CAN GO BEYOND 1099 00:43:58,280 --> 00:43:58,640 SPIKE. 1100 00:43:58,640 --> 00:44:00,200 EVERYBODY IS FOCUSING ON SPIKE. 1101 00:44:00,200 --> 00:44:02,800 LOOKING INTO WHAT IS THE 1102 00:44:02,800 --> 00:44:05,240 CONTRIBUTION OF THE SPIKE TO FOR 1103 00:44:05,240 --> 00:44:08,360 EXAMPLE VARIANT INFECTIVITY. 1104 00:44:08,360 --> 00:44:10,280 AND IMMUNE EVASION. 1105 00:44:10,280 --> 00:44:11,880 WE HAVE BEEN STARTING NOW TO 1106 00:44:11,880 --> 00:44:14,080 LOOK AT THE N M AND E PROTEINS, 1107 00:44:14,080 --> 00:44:15,200 TALKING ABOUT OUR FINDINGS IN 1108 00:44:15,200 --> 00:44:17,680 THE N PROTEIN TODAY, AS I SAID 1109 00:44:17,680 --> 00:44:20,160 THE N PROTEIN VERY IMPORTANT 1110 00:44:20,160 --> 00:44:22,280 PROTEIN, WE HAVE A LONG STANDING 1111 00:44:22,280 --> 00:44:23,640 INTEREST IN CAPSID NUCLEAR 1112 00:44:23,640 --> 00:44:26,600 CAPSID OR CORE PROTEINS IN OTHER 1113 00:44:26,600 --> 00:44:30,720 VIRUSES, THEY HAVE ENORMOUS HOST 1114 00:44:30,720 --> 00:44:32,720 BUT ALSO VIRALLY IMPORTANT 1115 00:44:32,720 --> 00:44:34,400 FUNCTIONS AND HERE THE NUCLEAR 1116 00:44:34,400 --> 00:44:38,920 CAPSID IS REALLY THE ONE THAT IS 1117 00:44:38,920 --> 00:44:41,800 MEDIATING RNA BINDING AND RNA 1118 00:44:41,800 --> 00:44:42,800 INCORPORATION TO ITS END 1119 00:44:42,800 --> 00:44:44,880 TERMINAL AND C TERMINAL DOMAIN 1120 00:44:44,880 --> 00:44:47,160 BUT ALSO OLIGOMERRIZES TO FORM 1121 00:44:47,160 --> 00:44:50,160 THE CAPSID AND THE PARTICLE OF 1122 00:44:50,160 --> 00:44:52,000 THE VIRUS. 1123 00:44:52,000 --> 00:44:53,720 IMPORTANTLY THE TWO N AND C 1124 00:44:53,720 --> 00:44:55,080 TERMINAL DOMAINS IS SEPARATED BY 1125 00:44:55,080 --> 00:44:57,640 A LINKER DOMAIN IN THE MIDDLE. 1126 00:44:57,640 --> 00:44:59,200 AND THIS LINKER DOMAIN CAUGHT 1127 00:44:59,200 --> 00:45:00,280 OUR ATTENTION BECAUSE MANY OF 1128 00:45:00,280 --> 00:45:05,560 THE MUTATIONS THAT HAVE BEEN 1129 00:45:05,560 --> 00:45:08,000 REPORTED FOR N PROTEINS IN 1130 00:45:08,000 --> 00:45:12,000 DIFFERENT VARIANTS HAVE ACTUALLY 1131 00:45:12,000 --> 00:45:13,280 CONCENTRATED IN THIS LINKER 1132 00:45:13,280 --> 00:45:13,760 REGION. 1133 00:45:13,760 --> 00:45:15,880 SO WHAT WE DID IN THIS PAPER IS 1134 00:45:15,880 --> 00:45:17,240 BASICALLY GENERATE N PROTEINS 1135 00:45:17,240 --> 00:45:20,880 WITH SIGNAL POINT MUTATIONS AND 1136 00:45:20,880 --> 00:45:23,400 ANY OF THESE THAT HAVE BEEN 1137 00:45:23,400 --> 00:45:25,640 REPORTED IN NUCLEAR CAPSID 1138 00:45:25,640 --> 00:45:27,440 PROTEINS AND WE FOUND THE ONES 1139 00:45:27,440 --> 00:45:31,480 MANY THE LINKER REGION 1140 00:45:31,480 --> 00:45:33,480 SPECIFICALLY HAVE HUGE INCREASE 1141 00:45:33,480 --> 00:45:35,720 IN INFECTIVITY WHEN YOU CAN DO 1142 00:45:35,720 --> 00:45:39,640 THE ASSAY I SHOWED YOU BEFORE. 1143 00:45:39,640 --> 00:45:44,160 THREE OF THEM, THE 9 -- 199L TO 1144 00:45:44,160 --> 00:45:46,040 WHICH WE ARE AND THE TWO OR 1145 00:45:46,040 --> 00:45:47,320 THREE M WHICH IS THE MUTATION 1146 00:45:47,320 --> 00:45:49,800 THAT IS FOUND IN DELTA VIRUS, 1147 00:45:49,800 --> 00:45:52,360 DELTA VARIANT, ACTUALLY VERY 1148 00:45:52,360 --> 00:45:54,120 MARKEDLY INCREASE INFECTIVITY UP 1149 00:45:54,120 --> 00:45:56,120 TO TENFOLD. 1150 00:45:56,120 --> 00:45:58,800 SO SYED LOOKED INTO WHAT EXACTLY 1151 00:45:58,800 --> 00:46:02,160 IS HAPPENING, WHY IS THERE MORE 1152 00:46:02,160 --> 00:46:04,200 LUCIFERASE IN THE TARGET CELLS 1153 00:46:04,200 --> 00:46:07,480 WHEN USES THESE MUTANTS. 1154 00:46:07,480 --> 00:46:10,080 HE LOOKED, PURIFIED THE 1155 00:46:10,080 --> 00:46:11,360 PARTICLES OF THE PRODUCTION, HE 1156 00:46:11,360 --> 00:46:13,240 LOOKED WHETHER THEY HAD MORE 1157 00:46:13,240 --> 00:46:14,960 SPIKE OR MORE PROCESS SPIKE. 1158 00:46:14,960 --> 00:46:19,120 THERE WAS NO CLEAR SIGNAL FOR N, 1159 00:46:19,120 --> 00:46:20,560 WE SAW TWO VARIANTS INCREASING 1160 00:46:20,560 --> 00:46:23,440 IN LEVELS BUT THE -- NOT FOR THE 1161 00:46:23,440 --> 00:46:25,760 THIRD WHICH WAS THE DELTA 1162 00:46:25,760 --> 00:46:27,600 VARIANT. 1163 00:46:27,600 --> 00:46:29,280 BUT THE MOST CONSISTENT SIGNAL 1164 00:46:29,280 --> 00:46:31,880 CAME FROM THE NORTHERN BLOT 1165 00:46:31,880 --> 00:46:33,480 WHERE HE COULD SHOW RNA 1166 00:46:33,480 --> 00:46:35,200 INCORPORATION IS ENHANCED IN ALL 1167 00:46:35,200 --> 00:46:36,120 THREE OF THEM. 1168 00:46:36,120 --> 00:46:38,880 THAT REALLY RESEMBLES THE 1169 00:46:38,880 --> 00:46:41,800 INCREASE IN INFECTIVITY THAT WE 1170 00:46:41,800 --> 00:46:48,400 SEE IN THE FUNCTIONAL ASSAY. 1171 00:46:48,400 --> 00:46:49,880 THE MODEL IS THESE M MUTATIONS 1172 00:46:49,880 --> 00:46:52,720 ARE INCREASING RNA 1173 00:46:52,720 --> 00:46:53,680 INCORPORATION, AND THEREFORE 1174 00:46:53,680 --> 00:46:57,080 INCREASE OR MAKING MORE 1175 00:46:57,080 --> 00:46:58,040 FUNCTIONAL VIRAL LIKE PARTICLES 1176 00:46:58,040 --> 00:47:01,840 IN THE SUPERNATANT, THAT LEADS 1177 00:47:01,840 --> 00:47:04,040 TO MORE INFECTION IN THE TARGET 1178 00:47:04,040 --> 00:47:04,480 CELL. 1179 00:47:04,480 --> 00:47:06,200 IN ORDER TO PROVE THIS IS REALLY 1180 00:47:06,200 --> 00:47:10,160 TRUE, WE NOW PUT THESE MUTATIONS 1181 00:47:10,160 --> 00:47:11,920 IN TO THE FULL LENGTH VIRUS. 1182 00:47:11,920 --> 00:47:14,920 WE USE INFECTIOUS CLONE 1183 00:47:14,920 --> 00:47:18,800 GENERATED BY PEI YONG SHI'S LAB 1184 00:47:18,800 --> 00:47:22,880 IN TEXAS, WE FOCUS ON THE S 2R 1185 00:47:22,880 --> 00:47:26,480 AND THE R 2 OR 3M MUTATION AND 1186 00:47:26,480 --> 00:47:28,920 YOU CAN SEE HERE INFECTIOUS 1187 00:47:28,920 --> 00:47:30,640 TITERS FROM THE SUPERNATANT OF 1188 00:47:30,640 --> 00:47:32,840 THESE TRANSFECTIONS THAT BOTH 1189 00:47:32,840 --> 00:47:35,800 MUTATIONS ARE REALLY INCREASING 1190 00:47:35,800 --> 00:47:38,000 INFECTIVITY OF THE VIRUSES TO A 1191 00:47:38,000 --> 00:47:39,800 SIMILAR DEGREE AS WE HAVE SEEN 1192 00:47:39,800 --> 00:47:42,040 IN THE BLPs. 1193 00:47:42,040 --> 00:47:44,800 THIS WAS ALSO TRUE FOR CLASSICAL 1194 00:47:44,800 --> 00:47:46,680 COMPETITION EXPERIMENTS WHERE WE 1195 00:47:46,680 --> 00:47:49,680 MIX THE MUTANT VIRUS WITH THE 1196 00:47:49,680 --> 00:47:51,560 WILD TYPE VIRUS AND WE CAN SEE 1197 00:47:51,560 --> 00:47:54,720 ALL THE DILUTIONS EVEN THE 1198 00:47:54,720 --> 00:47:56,440 HIGHEST DILUTION CAUGHT UP OR 1199 00:47:56,440 --> 00:47:57,840 WAS BETTER IN REPLICATING OR 1200 00:47:57,840 --> 00:48:00,160 CAUGHT UP WITH THE WILD TYPE IN 1201 00:48:00,160 --> 00:48:04,120 -- WITHIN TWO PASSAGES OF THE 1202 00:48:04,120 --> 00:48:05,640 VIRUSES. 1203 00:48:05,640 --> 00:48:07,080 INDICATING THESE HAVE INCREASE 1204 00:48:07,080 --> 00:48:08,960 INFECTIVITY AND REPLICATION 1205 00:48:08,960 --> 00:48:09,560 ADVANTAGES. 1206 00:48:09,560 --> 00:48:12,880 AS I SAID, THE MODEL IS THAT 1207 00:48:12,880 --> 00:48:14,880 THESE N MUTATIONS AND VARIANTS 1208 00:48:14,880 --> 00:48:16,440 ARE VERY CRITICAL. 1209 00:48:16,440 --> 00:48:19,080 FOR INCREASING INFECTIVITY AND 1210 00:48:19,080 --> 00:48:21,880 POTENTIAL TRANSMISSION OF THESE 1211 00:48:21,880 --> 00:48:22,440 VARIANTS. 1212 00:48:22,440 --> 00:48:25,320 BY ALLOWING THE VIRUS TO BE MORE 1213 00:48:25,320 --> 00:48:26,880 EFFECTIVE AND BUILD MORE 1214 00:48:26,880 --> 00:48:29,440 INFECTIOUS MORE EFFECTIVE MORE 1215 00:48:29,440 --> 00:48:30,960 EFFECTIVELY INFECTIOUS PARTICLES 1216 00:48:30,960 --> 00:48:33,200 THAT CAN MORE EFFECTIVELY INFECT 1217 00:48:33,200 --> 00:48:35,200 THE NEXT CELLS. 1218 00:48:35,200 --> 00:48:35,880 WE ARE CURRENTLY WORKING ON 1219 00:48:35,880 --> 00:48:37,080 THIS. 1220 00:48:37,080 --> 00:48:39,400 WE HAVE OF COURSE LIKE EVERYBODY 1221 00:48:39,400 --> 00:48:41,960 ELSE, ALSO FOCUSED OUR ATTENTION 1222 00:48:41,960 --> 00:48:46,280 ON THE NEW OMICRON VARIANT, 1223 00:48:46,280 --> 00:48:48,480 WHICH ALSO CONTAINS OF COURSE A 1224 00:48:48,480 --> 00:48:50,800 WHOLE RANGE OF SPIKE MUTATIONS 1225 00:48:50,800 --> 00:48:52,920 BUT ALSO CONTAINS A WHOLE SERIES 1226 00:48:52,920 --> 00:48:54,880 OF N MUTATIONS. 1227 00:48:54,880 --> 00:48:57,440 INCLUDING SOME IN THE LINKER 1228 00:48:57,440 --> 00:48:58,880 REGION I JUST POINTED OUT. 1229 00:48:58,880 --> 00:49:01,360 SO WHEN WE MADE A NUCLEAR CAPSID 1230 00:49:01,360 --> 00:49:03,280 THAT HAD ALL THE OMICRON 1231 00:49:03,280 --> 00:49:04,960 MUTATIONS AND COMPARED IT TO THE 1232 00:49:04,960 --> 00:49:07,480 NUCLEAR CAPSID, FROM DELTA, PUT 1233 00:49:07,480 --> 00:49:12,720 IT INTO VLPs, WE COULD SEE THE 1234 00:49:12,720 --> 00:49:14,040 OMICRON N PROTEIN IS MORE 1235 00:49:14,040 --> 00:49:16,320 EFFICIENT THAN THE DELTA TO 1236 00:49:16,320 --> 00:49:18,600 ENHANCE INFECTIVITY. 1237 00:49:18,600 --> 00:49:20,120 IF YOU JUST FOCUS ON TWO 1238 00:49:20,120 --> 00:49:21,680 MUTATIONS THAT ARE IN THE LINKER 1239 00:49:21,680 --> 00:49:24,560 REGION HERE WE SEE THAT THERE IS 1240 00:49:24,560 --> 00:49:26,040 ABOUT SIX FOLD INCREASE IN 1241 00:49:26,040 --> 00:49:28,360 INFECTIVITY BUT IT REALLY 1242 00:49:28,360 --> 00:49:30,880 REQUIRES THE WHOLE SLEW OF 1243 00:49:30,880 --> 00:49:31,960 OMICRON MUTATIONS HERE IN ORDER 1244 00:49:31,960 --> 00:49:34,080 TO ENHANCE DRAMATICALLY 1245 00:49:34,080 --> 00:49:36,440 INFECTIVITY OF PARTICLES THAT 1246 00:49:36,440 --> 00:49:38,480 CARRY OMICRON MY CLEAR CAPSID. 1247 00:49:38,480 --> 00:49:41,880 -- KNEW CLEAR CAP NUCLEAR 1248 00:49:41,880 --> 00:49:42,240 CAPSID. 1249 00:49:42,240 --> 00:49:44,200 SPIKE ALSO INCREASES 1250 00:49:44,200 --> 00:49:45,240 INFECTIVITY, IT IS A LITTLE BIT 1251 00:49:45,240 --> 00:49:46,240 DIFFERENT FROM WHAT WE OBSERVE 1252 00:49:46,240 --> 00:49:47,480 WITH DELTA. 1253 00:49:47,480 --> 00:49:49,760 BUT WE THINK THAT SPIKE AND 1254 00:49:49,760 --> 00:49:51,440 NUCLEAR CAPSID ARE SYNERGIZING 1255 00:49:51,440 --> 00:49:53,280 HERE TO MAKE THIS OMICRON VIRUS 1256 00:49:53,280 --> 00:49:55,080 VARIANT EXTREMELY INFECTIOUS. 1257 00:49:55,080 --> 00:49:57,080 THIS IS NOT THE CASE FOR THE M 1258 00:49:57,080 --> 00:50:00,080 AND E PROTEINS BECAUSE WHEN WE 1259 00:50:00,080 --> 00:50:02,560 PUT -- VERY RANDOM, WE HAVE M 1260 00:50:02,560 --> 00:50:06,480 AND E MUTATIONS IN VARIANTS BUT 1261 00:50:06,480 --> 00:50:09,440 OMICRON DOES, DELTA ALSO HAD N 1262 00:50:09,440 --> 00:50:11,120 MUTATION BUT YOU CAN SEE THAT 1263 00:50:11,120 --> 00:50:12,880 NEITHER OF THESE MUTATIONS IN 1264 00:50:12,880 --> 00:50:15,080 THE VARIANTS ACTUALLY ENHANCE 1265 00:50:15,080 --> 00:50:16,880 INFECTIVITY IN CONTRAST THEY 1266 00:50:16,880 --> 00:50:19,200 ACTUALLY DIMINISH THEM BUT 1267 00:50:19,200 --> 00:50:21,280 OBVIOUSLY THESE AFFECTS ARE 1268 00:50:21,280 --> 00:50:23,440 OVERRIDDEN BY THE POSITIVE 1269 00:50:23,440 --> 00:50:24,680 EFFECTS WE HAVE SEEN WITH 1270 00:50:24,680 --> 00:50:26,000 NUCLEAR CAPSID AND SPIKE BECAUSE 1271 00:50:26,000 --> 00:50:27,520 WHEN YOU LOOK AT THE FULL 1272 00:50:27,520 --> 00:50:29,000 PARTICLE THAT CONTAINS ALL OF 1273 00:50:29,000 --> 00:50:31,080 THE FOUR PROTEINS FROM OMICRON, 1274 00:50:31,080 --> 00:50:34,080 YOU CAN SEE THAT IT IS MORE 1275 00:50:34,080 --> 00:50:36,960 INFECTIOUS THAN DELTA VARIANT 1276 00:50:36,960 --> 00:50:38,120 HERE. 1277 00:50:38,120 --> 00:50:40,320 AND REALLY SHOWS THE SYNERGY OF 1278 00:50:40,320 --> 00:50:41,280 THE DIFFERENT MUTATIONS AT LEAST 1279 00:50:41,280 --> 00:50:43,040 IN THE STRUCTURAL PROTEINS FOR 1280 00:50:43,040 --> 00:50:44,160 THE VIRUS TO BECOME MORE 1281 00:50:44,160 --> 00:50:45,880 INFECTIOUS. 1282 00:50:45,880 --> 00:50:47,760 AS I SAID WE ARE VERY INTERESTED 1283 00:50:47,760 --> 00:50:48,880 IN THE N MUTATIONS. 1284 00:50:48,880 --> 00:50:51,680 WE ARE LOOKING VERY HARD INTO 1285 00:50:51,680 --> 00:50:53,480 HOW EXACTLY THESE N MUTATIONS 1286 00:50:53,480 --> 00:50:56,320 ARE ENHANCING RNA UPTAKE BECAUSE 1287 00:50:56,320 --> 00:50:57,680 THEY ARE IN THE LINKER REGION, 1288 00:50:57,680 --> 00:50:59,040 NOT NECESSARILY IN THE DOMAIN 1289 00:50:59,040 --> 00:51:00,320 NAYS ARE ATTACHING THE RNA 1290 00:51:00,320 --> 00:51:01,880 DIRECTLY. 1291 00:51:01,880 --> 00:51:03,880 IT IS VERY INTERESTING BECAUSE M 1292 00:51:03,880 --> 00:51:05,840 IS A HIGHLY PHOSPHORYLATED 1293 00:51:05,840 --> 00:51:07,200 PROTEIN AND PHOSPHORYLATION 1294 00:51:07,200 --> 00:51:09,000 SITES ARE IN THE LINKER REGIONS. 1295 00:51:09,000 --> 00:51:11,960 AND WE ARE TESTING DIFFERENT 1296 00:51:11,960 --> 00:51:13,840 ENZYMES THAT ARE KNOWN TO 1297 00:51:13,840 --> 00:51:15,080 PHOSPHORYLATE THIS REGION AND 1298 00:51:15,080 --> 00:51:16,520 SEEING HOW POTENTIALLY THE 1299 00:51:16,520 --> 00:51:17,640 PHOSPHORYLATION OF THIS PROTEIN 1300 00:51:17,640 --> 00:51:18,600 IS CHANGED. 1301 00:51:18,600 --> 00:51:22,480 IN THE VARIANTS AND HOW THAT 1302 00:51:22,480 --> 00:51:23,480 COULD EFFECT FUNCTION OF THE 1303 00:51:23,480 --> 00:51:25,120 NUCLEAR CAPSID. 1304 00:51:25,120 --> 00:51:27,120 OF COURSE THE VIRAL LIKE 1305 00:51:27,120 --> 00:51:30,200 PARTICLES ARE ALSO EXQUISITE, 1306 00:51:30,200 --> 00:51:34,280 SUBSTRATES TO TEST IMMUNE 1307 00:51:34,280 --> 00:51:34,880 FUNCTION. 1308 00:51:34,880 --> 00:51:38,920 SIMILAR TO PSEUDOTYPE VIRUS THAT 1309 00:51:38,920 --> 00:51:41,120 HAVE BEEN DONE QUITE A BIT FOR 1310 00:51:41,120 --> 00:51:42,840 OMICRON AND OTHER VIRUSES, WE 1311 00:51:42,840 --> 00:51:45,000 HAVE BEEN DEVELOPING VLP 1312 00:51:45,000 --> 00:51:45,920 NEUTRALIZATION ASSAYS. 1313 00:51:45,920 --> 00:51:49,000 WHERE WE JUST MEASURE THE 1314 00:51:49,000 --> 00:51:51,320 LUMINESCENCE AND THE ACCEPTOR 1315 00:51:51,320 --> 00:51:54,440 CELL AND DEVELOPING BASICALLY A 1316 00:51:54,440 --> 00:51:59,240 READ OF 50% INHIBITION HERE 1317 00:51:59,240 --> 00:52:01,240 NEUTRALIZATION, THAT WE USE AS 1318 00:52:01,240 --> 00:52:01,880 AN OUTREAD. 1319 00:52:01,880 --> 00:52:05,960 SO WITH THIS WE TESTED OMICRON 1320 00:52:05,960 --> 00:52:10,160 CARRYING BLPs. 1321 00:52:10,160 --> 00:52:11,520 AGAINST SERA OBTAINED FROM 1322 00:52:11,520 --> 00:52:14,280 PEOPLE WHO HAD OBTAINED TWO OF 1323 00:52:14,280 --> 00:52:19,200 THE PFIZER BIOTECH VACCINES, THE 1324 00:52:19,200 --> 00:52:21,520 MODERNA VACCINE OR J AND J 1325 00:52:21,520 --> 00:52:23,200 VACCINE OR RECOVERING FROM 1326 00:52:23,200 --> 00:52:25,320 PREVIOUS COVID INFECTIONS. 1327 00:52:25,320 --> 00:52:28,320 LIKE OTHERS WE HAVE OBSERVED 1328 00:52:28,320 --> 00:52:30,320 THAT OMICRON IS REALLY UNIQUELY 1329 00:52:30,320 --> 00:52:34,400 EVADING THE NEUTRALIZING 1330 00:52:34,400 --> 00:52:36,440 ACTIVITY IN THESE SERA BOTH FOR 1331 00:52:36,440 --> 00:52:40,800 THE mRNA VACCINES WITH SORT OF 1332 00:52:40,800 --> 00:52:42,600 SIMILAR REDUCTION WE OBSERVE 1333 00:52:42,600 --> 00:52:45,280 BETWEEN PFIZER AND MODERNA. 1334 00:52:45,280 --> 00:52:49,160 BUT WE REALLY SEE VERY LITTLE 1335 00:52:49,160 --> 00:52:51,280 NEUTRALIZING ACTIVITY WITH J AND 1336 00:52:51,280 --> 00:52:54,280 J VACCINE, THEY ALSO HAVE VERY 1337 00:52:54,280 --> 00:52:57,280 LITTLE TOTAL ANTI-SARS COV-2 1338 00:52:57,280 --> 00:52:59,720 ANTIBODIES IN THESE SERA AND IT 1339 00:52:59,720 --> 00:53:01,920 SPEAKS FOR POTENTIALLY THE P 1340 00:53:01,920 --> 00:53:05,880 CELL ACTIVATING ACTIVITY OF THIS 1341 00:53:05,880 --> 00:53:07,680 VACCINE TO EXPLAIN ITS 1342 00:53:07,680 --> 00:53:10,040 PROTECTION HERE. 1343 00:53:10,040 --> 00:53:12,880 WITH COVID, POST COVID SERA WE 1344 00:53:12,880 --> 00:53:14,120 INTERESTINGLY SAW DIFFERENT 1345 00:53:14,120 --> 00:53:14,760 RESULTS. 1346 00:53:14,760 --> 00:53:17,400 OMICRON VERY EFFECTIVE IN 1347 00:53:17,400 --> 00:53:19,080 EVADING THIS IMMUNITY WHILE FOR 1348 00:53:19,080 --> 00:53:23,880 DELTA WE SEE A WIDE RANGE OF 1349 00:53:23,880 --> 00:53:26,680 PROTECTIVE OR INHIBITORY RESULTS 1350 00:53:26,680 --> 00:53:27,160 HERE. 1351 00:53:27,160 --> 00:53:29,080 WE THINK THAT MOST PATIENTS HAVE 1352 00:53:29,080 --> 00:53:30,280 BEEN INFECTED WITH DELTA THAT 1353 00:53:30,280 --> 00:53:33,680 ARE INCLUDED HERE. 1354 00:53:33,680 --> 00:53:36,680 SO THE INFECTION WITH THE SAME 1355 00:53:36,680 --> 00:53:39,280 STRAIN MIGHT CONFER IN SOME 1356 00:53:39,280 --> 00:53:41,600 CASES QUITE EFFECTIVE IMMUNITY 1357 00:53:41,600 --> 00:53:42,280 HERE. 1358 00:53:42,280 --> 00:53:46,680 HOWEVER, THE BOOSTER WORKS IF WE 1359 00:53:46,680 --> 00:53:51,480 USE AND WE TEST SERA 16 DAYS OR 1360 00:53:51,480 --> 00:53:55,000 21 DAYS AFTER THE BOOST WE CAN 1361 00:53:55,000 --> 00:53:57,720 SEE THAT THE ANCESTRAL VIRUS 1362 00:53:57,720 --> 00:54:00,720 DELTA AND OMICRON SIGNIFICANTLY 1363 00:54:00,720 --> 00:54:03,120 REACTIONS ARE NEUTRALIZATION IS 1364 00:54:03,120 --> 00:54:06,080 SIGNIFICANTLY INCREASED. 1365 00:54:06,080 --> 00:54:08,200 AND BUT IF WE COMPARE THE 1366 00:54:08,200 --> 00:54:11,040 INCREASE BETWEEN ANCESTRAL DELTA 1367 00:54:11,040 --> 00:54:13,520 AND OMICRON WE SEE THE OMICRON 1368 00:54:13,520 --> 00:54:17,160 IS STILL ABLE TO ALSO SOMEHOW 1369 00:54:17,160 --> 00:54:19,400 EVADE THE NEUTRALIZING ACTIVITY 1370 00:54:19,400 --> 00:54:23,120 OF THE BOOST EVEN AFTER THREE 1371 00:54:23,120 --> 00:54:24,360 WEEKS AFTER THE BOOSTING. 1372 00:54:24,360 --> 00:54:27,320 SO THE BOOSTERS REALLY IS 1373 00:54:27,320 --> 00:54:30,320 WORKING BUT OMICRON IS STILL THE 1374 00:54:30,320 --> 00:54:34,160 ONE THAT CAN DEAL WITH IT THE 1375 00:54:34,160 --> 00:54:34,760 BEST. 1376 00:54:34,760 --> 00:54:37,080 WE ALSO TESTED THE AVAILABLE 1377 00:54:37,080 --> 00:54:39,080 MONOCLONAL ANTIBODIES FROM 1378 00:54:39,080 --> 00:54:39,560 REGENERON. 1379 00:54:39,560 --> 00:54:41,880 AS YOU KNOW THIS IS AN IC 50 1380 00:54:41,880 --> 00:54:43,440 EXPERIMENT SO HIGHER THE NUMBER 1381 00:54:43,440 --> 00:54:44,800 THE WORSE. 1382 00:54:44,800 --> 00:54:47,360 WE SEE THAT OMICRON REALLY HAS 1383 00:54:47,360 --> 00:54:48,680 ABSOLUTELY NO ACTIVITY AGAINST 1384 00:54:48,680 --> 00:54:50,200 THE TWO ANTIBODIES HERE. 1385 00:54:50,200 --> 00:54:51,960 WE ALSO GENERATED SOME SPIKE 1386 00:54:51,960 --> 00:54:53,680 PROTEINS THAT HAD MUTATIONS ONLY 1387 00:54:53,680 --> 00:54:57,840 IN THE CLASS 1 OR CLASS 3 SITES 1388 00:54:57,840 --> 00:55:00,600 AND THESE ANTIBODIES ARE AGAINST 1389 00:55:00,600 --> 00:55:03,160 CLASS 1 OR CLASS 3, YOU CAN SEE 1390 00:55:03,160 --> 00:55:05,400 THAT THEY ARE VERY EFFICIENT BUT 1391 00:55:05,400 --> 00:55:06,720 THAT THE COMBINATION OF THESE 1392 00:55:06,720 --> 00:55:15,360 MUTATIONS IS REALLY COMPLETELY 1393 00:55:15,360 --> 00:55:18,240 ABROGATING THE ACTIVITY OF THESE 1394 00:55:18,240 --> 00:55:20,040 IMPORTANT THERAPEUTICS. 1395 00:55:20,040 --> 00:55:23,000 SO AS A SUMMARY, I THINK THIS 1396 00:55:23,000 --> 00:55:25,040 SCURRY OF THE RNA PACKAGING 1397 00:55:25,040 --> 00:55:28,320 SIGNAL ALLOWS SARS COV-2 VLP 1398 00:55:28,320 --> 00:55:30,120 GENERAL RATION AS RAPID TOOL TO 1399 00:55:30,120 --> 00:55:32,480 ANALYZE IMMUNE EVASION AND 1400 00:55:32,480 --> 00:55:33,960 SPREAD OF THE BARS INFECTIVITY. 1401 00:55:33,960 --> 00:55:36,200 WE IDENTIFY SEVERAL N MUTATIONS 1402 00:55:36,200 --> 00:55:37,760 IN LINKER REGIONS THAT ENHANCE 1403 00:55:37,760 --> 00:55:40,880 VIRAL INFECTION BY ENHANCING RNA 1404 00:55:40,880 --> 00:55:41,280 PACKAGING. 1405 00:55:41,280 --> 00:55:42,960 AND THE RECOMBINANT VIRUS 1406 00:55:42,960 --> 00:55:44,920 CONFIRMED THIS, WE THINK VLPs 1407 00:55:44,920 --> 00:55:48,840 ARE RAPID TOOLS TO ANALYZE VIRUS 1408 00:55:48,840 --> 00:55:49,600 NEUTRALIZATION. 1409 00:55:49,600 --> 00:55:52,360 SORRY I REALIZE I'M A LITTLE 1410 00:55:52,360 --> 00:55:53,840 LATE BUT I WANT TO THANK 1411 00:55:53,840 --> 00:55:55,480 EVERYBODY I MENTIONED EVERYBODY 1412 00:55:55,480 --> 00:56:01,480 REALLY WANT TO THANK DAN, 1413 00:56:01,480 --> 00:56:03,480 JENNIFER, ANDREAS AND ALL OUR 1414 00:56:03,480 --> 00:56:04,480 COLLABORATORS AND FUNDERS HERE 1415 00:56:04,480 --> 00:56:06,360 FOR SUPPORTING US AND I'M HAPPY 1416 00:56:06,360 --> 00:56:08,280 TO TAKE -- THIS IS MY LAB, MY 1417 00:56:08,280 --> 00:56:10,520 WEB PAGE, IF YOU HAVE ANY MORE 1418 00:56:10,520 --> 00:56:11,240 QUESTIONS, HAPPY TO TAKE ANY 1419 00:56:11,240 --> 00:56:13,480 QUESTIONS. 1420 00:56:13,480 --> 00:56:17,440 >> THANK YOU, MELANIE, FOR 1421 00:56:17,440 --> 00:56:19,920 WONDERFUL TALK. 1422 00:56:19,920 --> 00:56:22,280 BEFORE I READ SOME OF THE 1423 00:56:22,280 --> 00:56:26,680 QUESTIONS, THE CME CODE FOR 1424 00:56:26,680 --> 00:56:29,280 TODAY IS 37961. 1425 00:56:29,280 --> 00:56:33,080 37961. 1426 00:56:33,080 --> 00:56:34,880 SO GOING BACK TO THE VERY 1427 00:56:34,880 --> 00:56:36,440 BEGINNING OF YOUR TALK, THERE 1428 00:56:36,440 --> 00:56:39,640 WERE A FEW QUESTIONS THAT I'M 1429 00:56:39,640 --> 00:56:42,200 GOING TO CONDENSE. 1430 00:56:42,200 --> 00:56:43,680 ONE IS IS THE RELIANCE OF THE 1431 00:56:43,680 --> 00:56:46,280 VIRUSES ON THE CHOLESTEROL 1432 00:56:46,280 --> 00:56:48,880 PATHWAY RELATED TO THE ENHANCED 1433 00:56:48,880 --> 00:56:51,200 VIRULENCE IN HUMANS WITH 1434 00:56:51,200 --> 00:56:51,720 OBESITY? 1435 00:56:51,720 --> 00:56:54,720 AND RELATED TO THIS, IS IT KNOWN 1436 00:56:54,720 --> 00:56:56,080 WHETHER BLOOD CHOLESTEROL LEVELS 1437 00:56:56,080 --> 00:56:57,960 ARE CORRELATED WITH COVID 1438 00:56:57,960 --> 00:56:59,640 INCIDENCE OR SEVERITY AND WHAT 1439 00:56:59,640 --> 00:57:00,840 ABOUT USE OF CHOLESTEROL 1440 00:57:00,840 --> 00:57:02,720 REDUCING DRUGS? 1441 00:57:02,720 --> 00:57:05,520 >> VERY GOOD QUESTIONS. 1442 00:57:05,520 --> 00:57:07,120 SO HERE, A FEW THINGS TO 1443 00:57:07,120 --> 00:57:08,720 CONSIDER. 1444 00:57:08,720 --> 00:57:13,000 IS THAT OBVIOUSLY OBESITY IS A 1445 00:57:13,000 --> 00:57:15,760 VERY STRONG RISK FACTOR. 1446 00:57:15,760 --> 00:57:20,160 WE HAVE STARTED TO MAKE FOR 1447 00:57:20,160 --> 00:57:21,880 SEVERE DISEASE AND WE ARE 1448 00:57:21,880 --> 00:57:22,680 LOOKING CAREFULLY INTO THIS. 1449 00:57:22,680 --> 00:57:25,560 I THINK WE HAVE TO DIFFERENTIATE 1450 00:57:25,560 --> 00:57:29,760 BETWEEN INFLAMMATORY EFFECTS OF 1451 00:57:29,760 --> 00:57:31,720 OBESITY AND ANTI-OBESE OR 1452 00:57:31,720 --> 00:57:32,880 ANTI-CHOLESTEROL MEDICATION AND 1453 00:57:32,880 --> 00:57:35,880 WE HAVE TO DIFFERENTIATE BETWEEN 1454 00:57:35,880 --> 00:57:37,960 DIRECT ANTIVIRAL OR ANTIVIRAL 1455 00:57:37,960 --> 00:57:39,440 EFFECTS. 1456 00:57:39,440 --> 00:57:43,520 SO WE THINK THAT THERE IS A 1457 00:57:43,520 --> 00:57:45,760 DIRECT LINK AND FOR EXAMPLE IF 1458 00:57:45,760 --> 00:57:48,280 WE FEED CELLS LBL IN VITRO WE 1459 00:57:48,280 --> 00:57:52,040 CAN INCREASE TITERS OF THE VIRUS 1460 00:57:52,040 --> 00:57:53,400 BY TWO LOG. 1461 00:57:53,400 --> 00:57:54,760 DEFINITELY THIS IS SOMETHING THE 1462 00:57:54,760 --> 00:57:59,680 VIRUS LIKES AS A PURE VIRAL 1463 00:57:59,680 --> 00:58:01,080 REPLICATION MACHINERY -- 1464 00:58:01,080 --> 00:58:01,640 MACHINE. 1465 00:58:01,640 --> 00:58:04,320 SO THAT IS SOMETHING THAT WE ARE 1466 00:58:04,320 --> 00:58:07,800 VERY MUCH INTERESTED IN. 1467 00:58:07,800 --> 00:58:09,720 IN TERMS OF ANTI-CHOLESTEROL 1468 00:58:09,720 --> 00:58:11,040 DRUGS, MANY PEOPLE HAVE LOOKED 1469 00:58:11,040 --> 00:58:14,120 INTO STATINS AND HAVE SHOWN THAT 1470 00:58:14,120 --> 00:58:16,440 STATINS HAVE SOME PROTECTIVE 1471 00:58:16,440 --> 00:58:18,880 EFFECT AGAINST SEVERE COVID. 1472 00:58:18,880 --> 00:58:21,120 BUT AGAIN, THE ANTI-INFLAMMATORY 1473 00:58:21,120 --> 00:58:24,080 EFFECTS HERE MIGHT BE VERY 1474 00:58:24,080 --> 00:58:24,320 IMPORTANT. 1475 00:58:24,320 --> 00:58:25,400 WHAT WE ARE INTERESTED IN IS 1476 00:58:25,400 --> 00:58:27,480 REALLY LOOKING AT DRUGS THAT 1477 00:58:27,480 --> 00:58:29,200 BLOCK INTRACELLULAR CHOLESTEROL 1478 00:58:29,200 --> 00:58:30,120 LEVELS BECAUSE WE THINK THAT 1479 00:58:30,120 --> 00:58:33,600 THAT IS WHAT THE VIRUS NEEDS AND 1480 00:58:33,600 --> 00:58:33,920 LIKES. 1481 00:58:33,920 --> 00:58:35,600 THE STATINS AND OTHER 1482 00:58:35,600 --> 00:58:36,840 ANTI-CHOLESTEROL DRUGS ARE 1483 00:58:36,840 --> 00:58:38,080 REALLY SHUFFLING THE CHOLESTEROL 1484 00:58:38,080 --> 00:58:39,760 INTO THE CELL. 1485 00:58:39,760 --> 00:58:41,880 WHICH MIGHT BE A DOUBLE EDGE 1486 00:58:41,880 --> 00:58:44,960 SWORD BECAUSE YOU MIGHT REDUCE 1487 00:58:44,960 --> 00:58:46,320 IT IN THE CIRCULATION BUT 1488 00:58:46,320 --> 00:58:47,600 INCREASE IN THE CELL. 1489 00:58:47,600 --> 00:58:49,440 SO THE DRUGS WE ARE USING 1490 00:58:49,440 --> 00:58:51,680 CURRENTLY ARE THOSE THAT ARE 1491 00:58:51,680 --> 00:58:54,840 GOING AGAINST INTRACELLULAR 1492 00:58:54,840 --> 00:58:57,960 PRODUCTION OF CHOLESTEROL AND 1493 00:58:57,960 --> 00:59:00,240 ARE HOPEFULLY LOWERING 1494 00:59:00,240 --> 00:59:01,480 INTRACELLULAR CHOLESTEROL LEVELS 1495 00:59:01,480 --> 00:59:05,440 AND DECREASING VIRAL REPLICATION 1496 00:59:05,440 --> 00:59:05,720 REPLICATION. 1497 00:59:05,720 --> 00:59:07,320 THIS IS SORT OF A LARGE 1498 00:59:07,320 --> 00:59:08,000 OVERVIEW, THERE IS AN 1499 00:59:08,000 --> 00:59:11,120 INTERESTING PAPER OUT FROM MY 1500 00:59:11,120 --> 00:59:11,720 COLLEAGUE KATHERINE BLISS AT 1501 00:59:11,720 --> 00:59:12,480 STANFORD WHO SHOWS THE VIRUS 1502 00:59:12,480 --> 00:59:14,920 DIRECTLY INFECTS ADIPOCYTES 1503 00:59:14,920 --> 00:59:16,200 WHICH IS SOMETHING THAT WE ARE 1504 00:59:16,200 --> 00:59:17,720 VERY INTERESTED IN AND WE ARE 1505 00:59:17,720 --> 00:59:19,640 TALKING CLOSELY WITH KATHERINE 1506 00:59:19,640 --> 00:59:22,960 TO SEE HOW WE CAN VERIFY THIS 1507 00:59:22,960 --> 00:59:24,880 AND POTENTIALLY USE THIS AS A 1508 00:59:24,880 --> 00:59:28,600 SPECIFIC STRATEGY. 1509 00:59:28,600 --> 00:59:31,040 >> HAVE YOUR STUDIES IDENTIFIED 1510 00:59:31,040 --> 00:59:33,920 THE MECHANISM OF WHY SARS COV-2 1511 00:59:33,920 --> 00:59:36,880 BUT NOT SEASONAL CORONA VIRUSES 1512 00:59:36,880 --> 00:59:40,400 LIKE LC 43 CAUSE CARDIAC 1513 00:59:40,400 --> 00:59:41,680 PATHOLOGY OR NEURAL PATHOLOGY? 1514 00:59:41,680 --> 00:59:44,920 >> YES, NO WE HAVEN'T BUT WE 1515 00:59:44,920 --> 00:59:46,720 WOULD LOVE, I THINK AS I SAID WE 1516 00:59:46,720 --> 00:59:49,160 ARE VERY INTERESTED IN THE 1517 00:59:49,160 --> 00:59:50,280 CARDIAC PATHOLOGY BECAUSE WE 1518 00:59:50,280 --> 00:59:52,280 HAVE BEEN WORKING VERY CLOSELY 1519 00:59:52,280 --> 00:59:56,280 THERE AND IS VERY FEW VIRAL 1520 00:59:56,280 --> 00:59:57,560 PARTICLES ARE NECESSARY TO 1521 00:59:57,560 --> 00:59:58,720 DAMAGE THE HEART. 1522 00:59:58,720 --> 01:00:00,440 SO THAT IS SOMETHING THAT WE ARE 1523 01:00:00,440 --> 01:00:02,160 DEFINITELY FOLLOWING UP WITH. 1524 01:00:02,160 --> 01:00:03,560 WE HAVE NOT DONE THE STUDIES 1525 01:00:03,560 --> 01:00:05,000 WITH THE COMMON COLD CORONA 1526 01:00:05,000 --> 01:00:06,120 VIRUSES HERE BUT I THINK WE 1527 01:00:06,120 --> 01:00:07,480 SHOULD. 1528 01:00:07,480 --> 01:00:09,320 THE OTHER THING IS THAT WE WORK 1529 01:00:09,320 --> 01:00:14,520 VERY CLOSELY WITH ARNOLD 1530 01:00:14,520 --> 01:00:16,600 CRICKSTEEN HERE AND ALSO WARNER 1531 01:00:16,600 --> 01:00:19,680 GREEN UCSF GLADSTONE LOOKING 1532 01:00:19,680 --> 01:00:24,320 WITH BRAIN ORGANOIDS INTO THE 1533 01:00:24,320 --> 01:00:27,000 BRAIN TRUMPISM AND ALSO 1534 01:00:27,000 --> 01:00:27,520 POTENTIAL DAMAGE. 1535 01:00:27,520 --> 01:00:29,280 I THINK THERE IS A HUGE AMOUNT 1536 01:00:29,280 --> 01:00:30,400 OF WORK TO BE DONE. 1537 01:00:30,400 --> 01:00:31,960 I THINK THE VIRUS IN OUR HANDS 1538 01:00:31,960 --> 01:00:35,240 REPLY CASE MOSTLY IN ASTROCYTES. 1539 01:00:35,240 --> 01:00:36,480 AND NOT IN NEURONS. 1540 01:00:36,480 --> 01:00:40,320 I THINK THERE IS NOW PAPERS OR 1541 01:00:40,320 --> 01:00:42,080 PRE-PRINT OUT SHOWING THAT 1542 01:00:42,080 --> 01:00:45,080 PEOPLE ACTUALLY RECOVER IN LONG 1543 01:00:45,080 --> 01:00:46,440 COVID INFECTIOUS PARTICLE FROM 1544 01:00:46,440 --> 01:00:48,680 THE BRAIN SO THERE MIGHT BE A 1545 01:00:48,680 --> 01:00:50,680 RESERVOIR FUNCTION HERE WHICH IS 1546 01:00:50,680 --> 01:00:54,240 SOMETHING WE DEFINITELY NEED TO 1547 01:00:54,240 --> 01:00:55,360 LOOK INTO. 1548 01:00:55,360 --> 01:00:57,680 ONE LAST THING, I THINK THE KEY 1549 01:00:57,680 --> 01:00:59,480 WITH COMMON COLD CORONA VIRUS IS 1550 01:00:59,480 --> 01:01:02,200 THEY REPLICATE TO MUCH LOWER 1551 01:01:02,200 --> 01:01:04,520 LEVELS THAN SARS COV-2, EITHER 1552 01:01:04,520 --> 01:01:07,520 WE HAVEN'T MADE IT INTO THE 1553 01:01:07,520 --> 01:01:08,440 RIGHT SYSTEM BUT DIRECT 1554 01:01:08,440 --> 01:01:09,360 COMPARISONS HERE ARE A LITTLE 1555 01:01:09,360 --> 01:01:13,280 BIT HAMPERED BY THE DIFFERENT 1556 01:01:13,280 --> 01:01:14,120 REPLICATION KINETICS OF THE 1557 01:01:14,120 --> 01:01:15,480 VIRUSES. 1558 01:01:15,480 --> 01:01:18,360 >> WE ARE GOING TO SWITCH TO 1559 01:01:18,360 --> 01:01:20,080 YOUR CRISPR BASED TESTING 1560 01:01:20,080 --> 01:01:22,640 APPARATUS. 1561 01:01:22,640 --> 01:01:24,400 CAN YOU KINETIC APPROACH 1562 01:01:24,400 --> 01:01:26,200 DISTINGUISH VARIANTS WITH ONE OR 1563 01:01:26,200 --> 01:01:26,960 TWO MISMATCHES IN DIFFERENT 1564 01:01:26,960 --> 01:01:27,760 POSITION? 1565 01:01:27,760 --> 01:01:29,760 >> YES, YES WE CAN. 1566 01:01:29,760 --> 01:01:31,360 I THINK IT IS STILL -- I MEAN, 1567 01:01:31,360 --> 01:01:33,080 WHAT MAKES A GUIDE WORK AND WHAT 1568 01:01:33,080 --> 01:01:35,480 MAKES A GUIDE NOT WORK IS STILL 1569 01:01:35,480 --> 01:01:37,720 SOME -- LITTLE BIT OF A MYSTERY. 1570 01:01:37,720 --> 01:01:41,080 I THINK THERE IS MANY ALGORITHMS 1571 01:01:41,080 --> 01:01:42,080 BEING WRITTEN ABOUT IT AND MANY 1572 01:01:42,080 --> 01:01:43,960 PEOPLE CLAIMED THEY KNOW HOW TO 1573 01:01:43,960 --> 01:01:46,520 DESIGN A GOOD GUIDE VERSUS A BAD 1574 01:01:46,520 --> 01:01:46,880 ONE. 1575 01:01:46,880 --> 01:01:49,240 IF THE MUTATION IS IN A CERTAIN 1576 01:01:49,240 --> 01:01:51,080 PLACE AND WE HAVE TO USE THAT 1577 01:01:51,080 --> 01:01:53,080 AREA BASICALLY IN ORDER TO 1578 01:01:53,080 --> 01:01:54,720 DESIGN THE GUIDE, I THINK WE ARE 1579 01:01:54,720 --> 01:01:59,480 STILL DEPENDENT ON CERTAIN 1580 01:01:59,480 --> 01:02:04,920 ASPECTS OF THE ACCESSIBILITY, 1581 01:02:04,920 --> 01:02:07,000 3-D FORMATION OF LOOPS AND STUFF 1582 01:02:07,000 --> 01:02:08,240 LIKE THIS WHICH WE HAVEN'T 1583 01:02:08,240 --> 01:02:09,680 REALLY FULLY MASTERED. 1584 01:02:09,680 --> 01:02:12,440 BUT I THINK IF WE HAVE IT IN AN 1585 01:02:12,440 --> 01:02:14,040 AREA THAT WORKS AS A GUIDE AND 1586 01:02:14,040 --> 01:02:16,920 WE CAN MOVE THE GUIDE UP AND 1587 01:02:16,920 --> 01:02:18,600 DOWN AND MOVE THE MUTATION IN 1588 01:02:18,600 --> 01:02:20,560 THE GUIDE UP AND DOWN, I THINK 1589 01:02:20,560 --> 01:02:24,480 WE HAVE VERY GOOD -- WE HAVE 1590 01:02:24,480 --> 01:02:26,560 VERY GOOD SENSIBILITY, 1591 01:02:26,560 --> 01:02:27,600 OPPORTUNITIES TO MAKE GUIDES 1592 01:02:27,600 --> 01:02:28,800 THAT CAN DIFFERENTIATE BETWEEN 1593 01:02:28,800 --> 01:02:31,000 SINGLE MUTANTS. 1594 01:02:31,000 --> 01:02:34,720 >> AND REGARDING THE LAST TOPIC 1595 01:02:34,720 --> 01:02:37,360 YOU DISCUSSED, HOW MUCH DO THE 1596 01:02:37,360 --> 01:02:38,680 MUTATIONS IN THE END CONTRIBUTE 1597 01:02:38,680 --> 01:02:41,520 TO MAKING THE OMICRON VIRUS MORE 1598 01:02:41,520 --> 01:02:41,880 VIRULENT? 1599 01:02:41,880 --> 01:02:46,360 CAN YOU USE YOUR SYSTEM TO LOOK 1600 01:02:46,360 --> 01:02:47,840 AT THE DIFFERENT CONTRIBUTIONS 1601 01:02:47,840 --> 01:02:50,880 OF THE SPIKE MUTATIONS VERSUS N 1602 01:02:50,880 --> 01:02:51,280 MUTATIONS? 1603 01:02:51,280 --> 01:02:53,640 >> WE FIND THE N MUTATIONS ARE 1604 01:02:53,640 --> 01:02:54,760 MORE IMPORTANT IN MAKING THE 1605 01:02:54,760 --> 01:02:55,480 VIRUS INFECTIOUS. 1606 01:02:55,480 --> 01:02:58,680 SO IF WE PUT AN OMICRON N INTO 1607 01:02:58,680 --> 01:03:00,480 AN VLP WE SEE A HIGHER 1608 01:03:00,480 --> 01:03:03,680 INFECTIVITY THAN IF WE PUT THE 1609 01:03:03,680 --> 01:03:05,480 SPIKE FROM OMICRON SPIKE ON VLP. 1610 01:03:05,480 --> 01:03:08,760 BUT WE THINK THAT BOTH TOGETHER 1611 01:03:08,760 --> 01:03:12,200 ARE REALLY MAKING THAT VIRUS 1612 01:03:12,200 --> 01:03:15,400 UNIQUELY MORE INFECTIOUS. 1613 01:03:15,400 --> 01:03:16,880 EVEN OVERRIDING THE NEGATIVE 1614 01:03:16,880 --> 01:03:18,840 EFFECTS OF THE M AND E MUTATIONS 1615 01:03:18,840 --> 01:03:20,760 THAT WE IDENTIFIED. 1616 01:03:20,760 --> 01:03:23,000 >> THE FINAL QUESTION, DO YOU 1617 01:03:23,000 --> 01:03:25,520 THINK THAT MUTATIONS IN THE N 1618 01:03:25,520 --> 01:03:27,840 PROTEIN VARIANTS ARE FOR 1619 01:03:27,840 --> 01:03:31,680 COMPETING OVER THE RNA BINDING 1620 01:03:31,680 --> 01:03:34,480 PROTEINS LIKE HOST SUPPRESSION 1621 01:03:34,480 --> 01:03:35,960 OF THE IMMUNE RESPONSE TO 1622 01:03:35,960 --> 01:03:37,040 SUPPRESS IMMUNE RESPONSE? 1623 01:03:37,040 --> 01:03:37,760 >> POSSIBLE. 1624 01:03:37,760 --> 01:03:38,560 VERY POSSIBLE AND WE DON'T KNOW 1625 01:03:38,560 --> 01:03:40,040 YET. 1626 01:03:40,040 --> 01:03:41,840 I THINK EVERYTHING IS ON THE 1627 01:03:41,840 --> 01:03:43,040 TABLE AT THIS POINT. 1628 01:03:43,040 --> 01:03:43,560 >> ALL RIGHT. 1629 01:03:43,560 --> 01:03:45,480 THANK YOU SO MUCH, MELANIE. 1630 01:03:45,480 --> 01:03:48,560 THIS ENDS TODAY'S LECTURE. 1631 01:03:48,560 --> 01:03:50,640 THANK YOU, EVERYONE. 1632 01:03:50,640 --> 01:07:08,400 >> THANK YOU, BYE-BYE.