1 00:00:05,720 --> 00:00:09,920 >>WELCOME TO THE 2 00:00:09,920 --> 00:00:11,800 MONTHLY DNA REPAIR INTEREST 3 00:00:11,800 --> 00:00:12,800 GROUP VIDEO CONFERENCES. 4 00:00:12,800 --> 00:00:15,560 TODAY WE ARE VERY GLAD TO HAVE 5 00:00:15,560 --> 00:00:20,000 DR. JO MORRIS JOIN US WHO IS 6 00:00:20,000 --> 00:00:27,480 FROM UNIVERSITY OF BIRMINGHAM. 7 00:00:27,480 --> 00:00:28,040 SHE STARTED IN INSTITUTE OF 8 00:00:28,040 --> 00:00:30,440 CANCER AND GENOMIC SCIENCES AND 9 00:00:30,440 --> 00:00:32,800 WORK AT KINGS COLLEGE IN LONDON 10 00:00:32,800 --> 00:00:34,120 WITH ED SOLOMON. 11 00:00:34,120 --> 00:00:37,080 BEFORE SETTING UP HER OWN GROUP 12 00:00:37,080 --> 00:00:42,520 AT UNITED -- UNIVERSITY AT 13 00:00:42,520 --> 00:00:47,720 BIRMINGHAM. 14 00:00:47,720 --> 00:00:50,960 SHE WAS ONE OF THE FIRST 15 00:00:50,960 --> 00:00:55,240 RESEARCHERS TO REPORT FOR 16 00:00:55,240 --> 00:00:58,800 MODIFICATION IN DNA DAMAGE 17 00:00:58,800 --> 00:01:06,520 RESPONSE AND INVESTIGATED 18 00:01:06,520 --> 00:01:12,200 BRC1LIGASE, THE ASSUME MONTH 19 00:01:12,200 --> 00:01:13,880 PROTEASE. 20 00:01:13,880 --> 00:01:17,640 DR. MORRIS HAS INVESTIGATED HOW 21 00:01:17,640 --> 00:01:21,480 BRC1 INTERACT WITH RD51 FOR 22 00:01:21,480 --> 00:01:22,640 REPLICATION FOR PROTECTION. 23 00:01:22,640 --> 00:01:26,560 SO WITHOUT FURTHER ADO WE WILL 24 00:01:26,560 --> 00:01:29,640 TRANSFER TO DR. MORRIS. 25 00:01:29,640 --> 00:01:31,240 FOR THE AUDIENCE IF YOU HAVE 26 00:01:31,240 --> 00:01:32,640 ANY QUESTIONS, PLEASE TYPE IT 27 00:01:32,640 --> 00:01:34,840 IN THE CHAT BOX. 28 00:01:34,840 --> 00:01:35,840 WE WILL GET TO THE QUESTIONS BY 29 00:01:35,840 --> 00:01:37,480 THE END OF THE TALK. 30 00:01:37,480 --> 00:01:38,240 DR. MORRIS? 31 00:01:38,240 --> 00:01:40,480 >>Jo Morris: CAN YOU HEAR ME? 32 00:01:40,480 --> 00:01:41,840 >>YES. 33 00:01:41,840 --> 00:01:45,320 >>Jo Morris: MY VIDEO CAMERA 34 00:01:45,320 --> 00:01:47,000 HAS JUST GONE DOWN, I WILL USE 35 00:01:47,000 --> 00:01:50,640 THE OTHER ONE, APOLOGIES FOR 36 00:01:50,640 --> 00:01:52,560 THE AWKWARD ANGLE. 37 00:01:52,560 --> 00:01:54,560 THANK YOU, ALL AND FOR INVITING 38 00:01:54,560 --> 00:01:57,000 ME SOME TIME AGO. 39 00:01:57,000 --> 00:01:58,880 >>WHAT I WOULD LIKE TO TALK TO 40 00:01:58,880 --> 00:02:00,160 YOU TODAY ABOUT IS THIS 41 00:02:00,160 --> 00:02:01,320 QUESTION HERE. 42 00:02:01,320 --> 00:02:04,280 WHAT UNDER LIES SYNTHETIC 43 00:02:04,280 --> 00:02:08,000 LETHAL RELATIONSHIP BETWEEN 44 00:02:08,000 --> 00:02:16,600 POLYMERASE THETA TARGETING AND 45 00:02:16,600 --> 00:02:18,280 BRCA 1/2 OR 53BP1 DEFICIENCY. 46 00:02:18,280 --> 00:02:21,720 THE ANSWER IS IT DEPENDS. 47 00:02:21,720 --> 00:02:25,960 WHICH DEFICIENCY AND HOW POL 48 00:02:25,960 --> 00:02:27,640 THETA IS TARGETED. 49 00:02:27,640 --> 00:02:29,480 MY APOLOGIES TO THOSE ON THE 50 00:02:29,480 --> 00:02:33,920 LINE WHO UNDERSTAND POL THETA 51 00:02:33,920 --> 00:02:36,040 BETTER THAN I, I APOLOGIZE. 52 00:02:36,040 --> 00:02:38,920 AND THEN TALK TO LOSS COMPARED 53 00:02:38,920 --> 00:02:39,800 TO ITS INHIBITION. 54 00:02:39,800 --> 00:02:41,920 AND I WILL TAKE A LITTLE DETOUR 55 00:02:41,920 --> 00:02:45,240 ALONG THE WAY TO TELL YOU ABOUT 56 00:02:45,240 --> 00:02:55,680 MODEL WHICH IS BRCA C61G. 57 00:03:08,240 --> 00:03:10,280 LET ME GET MY LASER POINTER 58 00:03:10,280 --> 00:03:11,200 BACK. 59 00:03:11,200 --> 00:03:16,040 WHERE IT FORMS MICRO FOLLOWED 60 00:03:16,040 --> 00:03:20,240 BY LIGATION TO REPAIR DNA 61 00:03:20,240 --> 00:03:23,320 BREAKS AFTER DISSECTION. 62 00:03:23,320 --> 00:03:24,360 THERE'S PAPERS PARTICULARLY 63 00:03:24,360 --> 00:03:26,360 THIS WEEK SUGGESTING A ROLE FOR 64 00:03:26,360 --> 00:03:35,920 THAT IS IN MITOSI. 65 00:03:35,920 --> 00:03:38,680 PRIOR TO THAT WE UNDERSTAND AS 66 00:03:38,680 --> 00:03:41,560 BACK UP PATHWAY. 67 00:03:41,560 --> 00:03:44,120 ONLY TO COMPLETE DNA REPAIR 68 00:03:44,120 --> 00:03:49,120 WHEN OTHER ASPECTS OF MOLECULAR 69 00:03:49,120 --> 00:03:55,800 COMBINATION WERE DISRUPTED. 70 00:03:55,800 --> 00:03:57,200 WHAT HAS BECOME PARTICULARLY 71 00:03:57,200 --> 00:03:59,800 INTERESTING IS THAT TARGETING 72 00:03:59,800 --> 00:04:04,200 POL THETA SEEMS TO BE SYNTHETIC 73 00:04:04,200 --> 00:04:05,800 LETHAL WITH A NUMBER OF 74 00:04:05,800 --> 00:04:06,520 DIFFERENT SETTINGS. 75 00:04:06,520 --> 00:04:10,880 THE MAIN INTERESTS HERE IS BRCA 76 00:04:10,880 --> 00:04:14,360 1 AND 2 DEFICIENCY BUT NOT THE 77 00:04:14,360 --> 00:04:16,280 ONLY DEFICIENCY SENSITIVE TO 78 00:04:16,280 --> 00:04:17,480 POL THETA. 79 00:04:17,480 --> 00:04:22,280 PAPERS HAVE SHOWN ANY NUMBER OF 80 00:04:22,280 --> 00:04:25,960 REPAIR DEFICIENCIES DEPEND ON 81 00:04:25,960 --> 00:04:29,760 POL THETA, ACTUALLY INCREASES 82 00:04:29,760 --> 00:04:30,760 SENSITIZATION. 83 00:04:30,760 --> 00:04:33,320 AND PERHAPS NOT SURPRISINGLY 84 00:04:33,320 --> 00:04:34,760 THEREFORE A NUMBER OF COMPANIES 85 00:04:34,760 --> 00:04:38,040 ARE INTERESTED IN GENERATING 86 00:04:38,040 --> 00:04:41,440 INHIBITORS TO THE POLYMERASE 87 00:04:41,440 --> 00:04:43,640 ACTIVITY AND SOME ARE SHOWN 88 00:04:43,640 --> 00:04:43,840 HERE. 89 00:04:43,840 --> 00:04:45,200 SO WE WANT TO UNDERSTAND A 90 00:04:45,200 --> 00:04:49,360 LITTLE BIT MORE ABOUT THIS. 91 00:04:49,360 --> 00:04:51,280 AT THE MOMENT, WHEN WE STARTED 92 00:04:51,280 --> 00:04:52,880 THIS PROJECT WHAT WAS 93 00:04:52,880 --> 00:04:58,160 UNDERSTOOD IS TARGETING POL 94 00:04:58,160 --> 00:05:00,200 THETA WAS WITH MORE RECEPTION 95 00:05:00,200 --> 00:05:03,480 AND REDUCE FILL IN OF NASCENT 96 00:05:03,480 --> 00:05:11,760 SIPGLE STRANDED GAPS AND 97 00:05:11,760 --> 00:05:13,080 INCREASE HR-INTERMEDIATES. 98 00:05:13,080 --> 00:05:21,120 SUGGESTION IT IS SIGNIFICANT IN 99 00:05:21,120 --> 00:05:22,920 BRCA 2 DEFICIENT. 100 00:05:22,920 --> 00:05:32,000 WE ARE USING BRCA ZYGOTE RAT 101 00:05:32,000 --> 00:05:35,280 CELLS. 102 00:05:35,280 --> 00:05:39,040 THEY DIE IN REGENESIS AND IN 103 00:05:39,040 --> 00:05:42,760 COMMON WITH OTHER MUTANT MODELS 104 00:05:42,760 --> 00:05:45,600 IF WE COMBINE THEM WITH 105 00:05:45,600 --> 00:05:49,280 DEFICIENT ANIMALS WE CAN RESCUE 106 00:05:49,280 --> 00:05:51,840 THAT LETHALITY. 107 00:05:51,840 --> 00:05:53,720 WE HAVE LIVE ANIMALS. 108 00:05:53,720 --> 00:05:59,520 THAT ALLOWS US TO LOOK AT THE 109 00:05:59,520 --> 00:06:04,640 BRCA DEFICIENCY ON THE BACK OF 110 00:06:04,640 --> 00:06:08,040 BP1. 111 00:06:08,040 --> 00:06:10,040 THEY HAVE RESISTANT. 112 00:06:10,040 --> 00:06:11,480 FEATURES YOU NORMALLY CONSIDER 113 00:06:11,480 --> 00:06:16,400 TO BE INDICATORS OF HR 114 00:06:16,400 --> 00:06:20,000 COMPETENCY BUT INTRIGUINGLY 115 00:06:20,000 --> 00:06:22,040 NEVERTHELESS SENSITIVE, THEY 116 00:06:22,040 --> 00:06:24,880 ARE HR COMPETENT YET SENSITIVE, 117 00:06:24,880 --> 00:06:27,680 SORT OF THE BASIS OF THE 118 00:06:27,680 --> 00:06:29,080 BEGINNING OF THIS INVESTIGATION. 119 00:06:29,080 --> 00:06:31,600 IF YOU TAKE A LOOK AT THESE 120 00:06:31,600 --> 00:06:34,440 CELLS, RADIATE AND ASK WHERE 121 00:06:34,440 --> 00:06:37,240 BRCA 1 IS, IT'S CLEAR THEY ARE 122 00:06:37,240 --> 00:06:40,880 NOT ENTIRELY NORMAL, IN TERMS 123 00:06:40,880 --> 00:06:43,280 OF THEIR BRCA -- THEY HAVE 124 00:06:43,280 --> 00:06:48,000 ABOUT 30% WHAT YOU WOULD EXPECT. 125 00:06:48,000 --> 00:06:49,960 20% OF WHAT YOU WOULD EXPECT. 126 00:06:49,960 --> 00:06:53,600 THAT LED US TO WONDER DESPITE 127 00:06:53,600 --> 00:06:55,920 THE HR COMPETENCY IF THEY ARE 128 00:06:55,920 --> 00:07:00,720 NOT USING THE PATHWAY TO 129 00:07:00,720 --> 00:07:11,280 SUPPORT THE HOMOLOGOUS -- WE 130 00:07:12,560 --> 00:07:17,200 KNOW BRCA BARD1, THE LOADING OF 131 00:07:17,200 --> 00:07:21,200 RAD51 CELLS DEFICIENCY FOR BRCA 132 00:07:21,200 --> 00:07:23,960 1, 168 INDEPENDENTLY AND 133 00:07:23,960 --> 00:07:26,960 ENABLES FOR RAD51 RECRUITMENT 134 00:07:26,960 --> 00:07:33,560 AND ROLL FOR 52 FOR BOTH 135 00:07:33,560 --> 00:07:36,320 IMPROVING -- SEPARATELY. 136 00:07:36,320 --> 00:07:38,680 THESE ARE INVOLVED IN THESE 137 00:07:38,680 --> 00:07:41,480 MUTANTS, LET'S SAY SLIGHTLY 138 00:07:41,480 --> 00:07:41,880 COMPROMISED CELLS. 139 00:07:41,880 --> 00:07:43,600 SO WE USED A NUMBER OF ASSAYS 140 00:07:43,600 --> 00:07:49,240 BUT THE ONE I WILL SHOW YOU IS 141 00:07:49,240 --> 00:07:50,960 CRISPR CAS9. 142 00:07:50,960 --> 00:07:52,600 ALLOWS US TO MEASURE THE AMOUNT 143 00:07:52,600 --> 00:07:54,320 OF HR. 144 00:07:54,320 --> 00:07:56,400 AND PERFORM THESE ASSAYS IN 145 00:07:56,400 --> 00:07:58,480 THESE CELLS AND WHERE WHETHER 146 00:07:58,480 --> 00:08:02,280 THEY ARE INDEED DEPENDENT ON HR 147 00:08:02,280 --> 00:08:02,720 SUPPORT MECHANISMS. 148 00:08:02,720 --> 00:08:04,840 THEY REQUIRE THE MUTANTS, BRCA 149 00:08:04,840 --> 00:08:07,680 1, IF WE DEPLETE WE SEE 150 00:08:07,680 --> 00:08:10,280 INCREASE IN HR BUT ALSO 151 00:08:10,280 --> 00:08:13,320 DEPENDENT ON 168 AND INDEED 152 00:08:13,320 --> 00:08:15,080 BRCA 152. 153 00:08:15,080 --> 00:08:18,760 IF HR IS SUPPORTED BY NON 154 00:08:18,760 --> 00:08:20,800 CONICAL MEANS. 155 00:08:20,800 --> 00:08:22,440 WE WONDERED IF THIS SUPPORT HAD 156 00:08:22,440 --> 00:08:25,320 SOMETHING TO DO WITH THE POL 157 00:08:25,320 --> 00:08:27,840 THETA SENSITIVITY. 158 00:08:27,840 --> 00:08:32,040 AND THE PUNCHLINE IS IT DOES. 159 00:08:32,040 --> 00:08:33,440 WE DEPLETED RF168 AND DIDN'T 160 00:08:33,440 --> 00:08:35,760 SEE ANYTHING BUT SEE SOMETHING 161 00:08:35,760 --> 00:08:38,560 DIFFERENT WITH THE 152. 162 00:08:38,560 --> 00:08:41,440 WE TREAT THESE CELLS, WE SEE 163 00:08:41,440 --> 00:08:42,120 REDUCTION IN VIABILITY. 164 00:08:42,120 --> 00:08:48,640 BUT WHEN WE TREAT THEM WITH 165 00:08:48,640 --> 00:08:52,120 BRCA152 INHIBITOR WHICH 166 00:08:52,120 --> 00:08:54,680 DISRUPTS, WE SEE THE SAME 167 00:08:54,680 --> 00:08:56,760 THING, THIS INTRIGUING BELL 168 00:08:56,760 --> 00:09:00,240 CURVE WHERE THERE IS A SWEET 169 00:09:00,240 --> 00:09:07,840 SPOT OF RAD 52 INHIBITOR. 170 00:09:07,840 --> 00:09:11,800 RESCUE OF THESE CELLS FOR POL 171 00:09:11,800 --> 00:09:12,680 THETA. 172 00:09:12,680 --> 00:09:14,760 WE SEE THE SAME THING WHEN WE 173 00:09:14,760 --> 00:09:17,080 TAKE HUMAN CELLS AND DEPLETE 174 00:09:17,080 --> 00:09:18,720 THEM WITH BRCA 1. 175 00:09:18,720 --> 00:09:22,720 SO WHAT'S GOING ON? 176 00:09:22,720 --> 00:09:26,880 THE AMOUNTS OF RAD52 IS NOT 177 00:09:26,880 --> 00:09:28,960 ENOUGH TO RESULT IN SENSITIVITY 178 00:09:28,960 --> 00:09:31,560 BUT IF WE USE A SIGNIFICANT 179 00:09:31,560 --> 00:09:32,840 AMOUNT MORE, WE DO SEE 180 00:09:32,840 --> 00:09:33,440 SENSITIVITY. 181 00:09:33,440 --> 00:09:35,360 THE AMOUNT WE ARE USING IS 182 00:09:35,360 --> 00:09:40,240 QUITE LOW. 183 00:09:40,240 --> 00:09:43,960 RAD52 ITSELF WE SEE IS 184 00:09:43,960 --> 00:09:52,600 RECRUITED TO FO SI IN CELLS BUT 185 00:09:52,600 --> 00:09:56,040 MORE FOR DOUBLE MUTANT -- 186 00:09:56,040 --> 00:10:03,960 WE KNOW THAT RAD52 INTERACTS 187 00:10:03,960 --> 00:10:05,600 WITH, IS THERE SOMEONE ELSE WHO 188 00:10:05,600 --> 00:10:07,440 CAN'T SEE THE SLIDES? 189 00:10:07,440 --> 00:10:08,960 IS THERE ANYTHING YOU CAN DO 190 00:10:08,960 --> 00:10:11,280 ABOUT THAT? 191 00:10:11,280 --> 00:10:14,440 NO? 192 00:10:14,440 --> 00:10:18,120 >> QUESTION FOR YOU. 193 00:10:18,120 --> 00:10:20,240 I'M SEEING A BOX ITSELF. 194 00:10:20,240 --> 00:10:21,960 IS THERE ANYTHING HOVERING OVER 195 00:10:21,960 --> 00:10:23,160 YOUR SLIDES? 196 00:10:23,160 --> 00:10:27,320 LIKE ANOTHER MENU OR? 197 00:10:27,320 --> 00:10:28,920 >>Karen Usdin: JEROME IN 198 00:10:28,920 --> 00:10:30,720 ADDITION TO THAT, IT SEEMS SOME 199 00:10:30,720 --> 00:10:31,920 AREN'T ABLE TO SEE THE SLIDES 200 00:10:31,920 --> 00:10:36,080 AT ALL. 201 00:10:36,080 --> 00:10:38,400 >>Chun Zhang Yang: STOP 202 00:10:38,400 --> 00:10:41,600 SHARING FOR ONE MINUTE AND THEN 203 00:10:41,600 --> 00:10:42,600 START SHARING THE APPLICATION 204 00:10:42,600 --> 00:10:50,520 AGAIN. 205 00:10:50,520 --> 00:10:52,440 >>Jo Morris: HAVE I STOPPED 206 00:10:52,440 --> 00:10:52,920 SHARING? 207 00:10:52,920 --> 00:10:56,160 >>Chun Zhang Yang: NOT YET. 208 00:10:59,160 --> 00:11:00,280 >>Jerome Battle: GO THROUGH 209 00:11:00,280 --> 00:11:01,520 THE SAME PROCESS YOU WENT 210 00:11:01,520 --> 00:11:10,200 THROUGH YESTERDAY. 211 00:11:10,200 --> 00:11:10,960 HERE YOU ARE. 212 00:11:10,960 --> 00:11:17,520 SHARE. 213 00:11:17,520 --> 00:11:19,080 >>Jo Morris: IT SAYS IT'S 214 00:11:19,080 --> 00:11:21,440 BLOCKED. 215 00:11:21,440 --> 00:11:24,360 >>Karen Usdin: WE CAN SEE YOUR 216 00:11:24,360 --> 00:11:27,480 SLIDES THOUGH THERE'S STILL 217 00:11:27,480 --> 00:11:29,080 GRAY BOXES BLOCKING, THE TOP 218 00:11:29,080 --> 00:11:30,680 AND BOTTOM, THE PERSON WHO 219 00:11:30,680 --> 00:11:33,800 COULDN'T SEE IT PREVIOUSLY CAN 220 00:11:33,800 --> 00:11:35,440 SEE IT NOW, SO AT LEAST THAT'S 221 00:11:35,440 --> 00:11:36,160 CHANGED. 222 00:11:36,160 --> 00:11:37,560 IT LOOKS LIKE SOMETIMES THE 223 00:11:37,560 --> 00:11:39,200 TITLE OF YOUR SLIDE IS BLOCKED 224 00:11:39,200 --> 00:11:40,000 OUT A BIT. 225 00:11:40,000 --> 00:11:42,000 IF YOU CAN REPEAT THE TITLE OF 226 00:11:42,000 --> 00:11:43,760 THE SLIDE AS YOU MOVE THROUGH 227 00:11:43,760 --> 00:11:43,960 IT? 228 00:11:43,960 --> 00:11:45,760 >>Jo Morris: OKAY, OKAY. 229 00:11:45,760 --> 00:11:48,360 ALL RIGHT. 230 00:11:48,360 --> 00:11:52,040 WHERE WERE WE? 231 00:11:52,040 --> 00:11:53,720 >>Jerome Battle: GO AHEAD AND 232 00:11:53,720 --> 00:11:54,840 PUT IT IN PRESENTATION MODE. 233 00:11:54,840 --> 00:12:00,200 >>Jo Morris: THANK YOU. 234 00:12:00,200 --> 00:12:02,520 THERE? 235 00:12:02,520 --> 00:12:04,040 >>Karen Usdin: SOME OF US CAN 236 00:12:04,040 --> 00:12:06,440 SEE IT. 237 00:12:06,440 --> 00:12:07,600 WE HAVEN'T HEARD WHETHER THE 238 00:12:07,600 --> 00:12:08,920 PEOPLE HAVING TROUBLE ARE STILL 239 00:12:08,920 --> 00:12:10,120 HAVING TROUBLE. 240 00:12:10,120 --> 00:12:10,920 THE TITLE IS BLOCKED. 241 00:12:10,920 --> 00:12:12,760 IF YOU CAN READ OUT THE TITLE 242 00:12:12,760 --> 00:12:15,560 OF EACH SLIDE. 243 00:12:15,560 --> 00:12:20,280 >>Jo Morris: THE APPLICATION, 244 00:12:20,280 --> 00:12:21,200 THAT'S YOU. 245 00:12:21,200 --> 00:12:23,040 THE WEB THING. 246 00:12:23,040 --> 00:12:24,760 >>Jerome Battle: OKAY, SO 247 00:12:24,760 --> 00:12:27,800 CLICK ANYWHERE IN AN OPEN SPACE 248 00:12:27,800 --> 00:12:30,440 SO THAT, IT SHOULD GO AWAY. 249 00:12:30,440 --> 00:12:32,240 >>Jo Morris: NO, IT DOESN'T, 250 00:12:32,240 --> 00:12:36,280 IT STAYS. 251 00:12:36,280 --> 00:12:39,320 >>Karen Usdin: OKAY, SHOULD WE 252 00:12:39,320 --> 00:12:40,160 JUST PROCEED. 253 00:12:40,160 --> 00:12:41,880 IF YOU JUST READ THE TITLE OF 254 00:12:41,880 --> 00:12:43,360 THE SLIDE IT SHOULD STILL BE 255 00:12:43,360 --> 00:12:43,920 OKAY. 256 00:12:43,920 --> 00:12:46,760 >>Jo Morris: I OF COURSE CAN'T 257 00:12:46,760 --> 00:12:50,360 SEE THE TITLE, OKAY, WHERE WERE 258 00:12:50,360 --> 00:12:50,560 WE? 259 00:12:50,560 --> 00:12:54,600 I HAVE TOLD YOU THE DOUBLE 260 00:12:54,600 --> 00:12:56,520 MUTANT CELLS ARE SENSITIVE AND 261 00:12:56,520 --> 00:13:00,240 SEE INCREASE IN RAD52 WHEN WE 262 00:13:00,240 --> 00:13:02,960 REMOVE POL THETA, WE CAN REMOVE 263 00:13:02,960 --> 00:13:07,440 THE SENSITIVITY BY SLIGHTLY 264 00:13:07,440 --> 00:13:10,280 DEPLETING AND DEPLETING RAD 52. 265 00:13:10,280 --> 00:13:11,920 IT INTERACTS WITH SINGLE 266 00:13:11,920 --> 00:13:14,240 STRANDED DNA IN CORPORATION 267 00:13:14,240 --> 00:13:16,320 WITH SINGLE STRANDED BINDED 268 00:13:16,320 --> 00:13:16,960 PROTEIN. 269 00:13:16,960 --> 00:13:18,960 I WONDERED IF WE COULD PERFORM 270 00:13:18,960 --> 00:13:24,440 THE SAME TRICK OF RESTORING 271 00:13:24,440 --> 00:13:29,600 SURVIVAL BY SLIGHTLY REPEATING 272 00:13:29,600 --> 00:13:29,880 RPA ITSELF. 273 00:13:29,880 --> 00:13:30,960 WE TRIED THAT. 274 00:13:30,960 --> 00:13:34,800 YOU CAN SEE LOW-LEVEL RPA 275 00:13:34,800 --> 00:13:35,040 DEPLETION. 276 00:13:35,040 --> 00:13:37,720 WHICH IS INDEED CAPABLE OF 277 00:13:37,720 --> 00:13:41,560 REDUCING RAD 52 FOR THOSE 278 00:13:41,560 --> 00:13:46,000 CELLS, DEPLETED FOR SIRPA, 279 00:13:46,000 --> 00:13:52,960 SORRY, POL THETA. 280 00:13:52,960 --> 00:13:56,160 WE SEE LESS RAD 52 IF WE 281 00:13:56,160 --> 00:13:59,240 DEPLETE RPA AND SIMILARLY LOW 282 00:13:59,240 --> 00:14:02,080 LEVEL CAN RESTORE SURVIVAL OF 283 00:14:02,080 --> 00:14:07,840 CELLS EXPOSED TO POL THETA SIRPA 284 00:14:07,840 --> 00:14:11,080 RAD 52 IS RESPONSIBLE FOR THE 285 00:14:11,080 --> 00:14:14,120 LETHALITY OF CELLS EXPOSED TO 286 00:14:14,120 --> 00:14:16,960 THE POL THETA SIRPA. 287 00:14:16,960 --> 00:14:21,320 TO CHECK THIS MORE CAREFULLY WE 288 00:14:21,320 --> 00:14:26,760 KNOW RPA IS CAPABLE OF CREATING 289 00:14:26,760 --> 00:14:28,960 PROTEASES WE PRODUCED AND ONE 290 00:14:28,960 --> 00:14:31,640 THAT LACKS THE BINDING REGION 291 00:14:31,640 --> 00:14:33,800 WHICH DOES STOP WITH 292 00:14:33,800 --> 00:14:36,000 INTERACTING WITH RPA. 293 00:14:36,000 --> 00:14:40,080 SO IF WE DEPLETE RAD52 SLIGHTLY 294 00:14:40,080 --> 00:14:50,200 WITH LOW CONCENTRATION of si 295 00:14:50,200 --> 00:14:55,320 RPA. 296 00:14:55,320 --> 00:14:59,120 IF WE EXPRESS SIRPA RESIST 297 00:14:59,120 --> 00:15:00,960 ANESTHE ARE MORE SENSITIVE TO 298 00:15:00,960 --> 00:15:03,960 THE POL THETA LOSS, IT IS THE 299 00:15:03,960 --> 00:15:06,320 RAD 52 RESPONSIBLE FOR 300 00:15:06,320 --> 00:15:08,840 SENSITIVITY OF POL THETA LOSS. 301 00:15:08,840 --> 00:15:11,440 WHEREAS IF WE GIVE RAD 52 302 00:15:11,440 --> 00:15:14,440 LACKING THE ABILITY TO INTERACT 303 00:15:14,440 --> 00:15:16,440 WITH RPA WE DON'T SEE THAT. 304 00:15:16,440 --> 00:15:18,920 SUGGESTING IT'S THE RAD 52 THAT 305 00:15:18,920 --> 00:15:20,480 CONTRIBUTES TO THE SENSITIVITY 306 00:15:20,480 --> 00:15:21,880 OF POL THETA LOSS. 307 00:15:21,880 --> 00:15:24,960 IT WASN'T A SURPRISE TO US IT'S 308 00:15:24,960 --> 00:15:27,560 RPA RAD 52 THAT APPEARS TO BE 309 00:15:27,560 --> 00:15:29,480 RESPONSIBLE TO THE SENSITIVITY 310 00:15:29,480 --> 00:15:32,560 OF THESE CELLS FOR THE REMOVAL 311 00:15:32,560 --> 00:15:33,680 OF POL THETA. 312 00:15:33,680 --> 00:15:36,520 WE HAVE KNOWN FOR A WHILE THAT 313 00:15:36,520 --> 00:15:38,600 RESECTION IS ASSOCIATED WITH 314 00:15:38,600 --> 00:15:40,200 SENSITIVITY TO POL THETA 315 00:15:40,200 --> 00:15:41,400 TARGETING. 316 00:15:41,400 --> 00:15:43,600 WE ASSUMED, AS IT TURNED OUT 317 00:15:43,600 --> 00:15:46,840 INCORRECTLY, IT MIGHT BE THE 318 00:15:46,840 --> 00:15:54,680 CASE OF RPAR52 -- WE TREATED 319 00:15:54,680 --> 00:15:59,200 CELLS WITH MRE 11 EXO NUCLEASE 320 00:15:59,200 --> 00:16:03,400 AS WE SAW LOW LEVEL, LOW 321 00:16:03,400 --> 00:16:04,560 CONCENTRATION TREATMENT RESCUE 322 00:16:04,560 --> 00:16:10,800 THE SURVIVAL OF THESE CELLS TO 323 00:16:10,800 --> 00:16:12,680 POL THETA SIRPA. 324 00:16:12,680 --> 00:16:16,440 BUT IT DID NOTHING, IT APPEARS 325 00:16:16,440 --> 00:16:22,640 RAD 52 IS NOT DOWNSTREAM OF MRE 326 00:16:22,640 --> 00:16:25,480 11, AT LEAST NOT WITH THESE 327 00:16:25,480 --> 00:16:26,640 CONCENTRATIONS TO SUPPRESS MRE 328 00:16:26,640 --> 00:16:27,560 11. 329 00:16:27,560 --> 00:16:30,120 WHICH LED US TO THE POSSIBILITY 330 00:16:30,120 --> 00:16:34,320 ACTUALLY MRE 11 IS DOWNSTREAM 331 00:16:34,320 --> 00:16:35,720 OF RAD52 RPA SUGGESTED BY A 332 00:16:35,720 --> 00:16:38,160 PAPER A FEW YEARS AGO FROM 333 00:16:38,160 --> 00:16:39,200 LOPEZ'S GROUP. 334 00:16:39,200 --> 00:16:45,080 WE DID AN ASSAY WE MEASURED THE 335 00:16:45,080 --> 00:16:47,400 SINGLE STRANDED DNA EXPOSED TO 336 00:16:47,400 --> 00:16:49,080 THE POL THETA TREATMENT AND HOW 337 00:16:49,080 --> 00:16:52,600 MUCH WAS ASSOCIATED WITH MRE 11. 338 00:16:52,600 --> 00:16:55,840 SO THIS IS THE ASSAY, WE LOOK 339 00:16:55,840 --> 00:16:59,360 AT MRE 11 ASSOCIATED WITH THE 340 00:16:59,360 --> 00:17:02,440 EXPOSED RDAU AFTER GIVEN FOR 24 341 00:17:02,440 --> 00:17:05,960 HOURS AND ANTIBODY CAPABLE OF 342 00:17:05,960 --> 00:17:08,320 PICKING IT UP IN SINGLE 343 00:17:08,320 --> 00:17:09,840 STRANDED. 344 00:17:09,840 --> 00:17:12,200 THERE'S MRE 11 ASSOCIATED WITH 345 00:17:12,200 --> 00:17:14,760 THAT SINGLE STRANDED DNA. 346 00:17:14,760 --> 00:17:17,240 INTRIGUINGLY WHEN WE GIVE LOW 347 00:17:17,240 --> 00:17:20,040 LEVEL CONCENTRATION OF RAD 52 348 00:17:20,040 --> 00:17:23,240 INHIBITOR THAT IS DECREASED 349 00:17:23,240 --> 00:17:23,560 DRASTICALLY. 350 00:17:23,560 --> 00:17:25,840 IT WOULD AMEER MRE 11 IS 351 00:17:25,840 --> 00:17:28,160 DOWNSTREAM OF RAD 52 RPA, 352 00:17:28,160 --> 00:17:31,360 SOMETHING LIKE THAT. 353 00:17:31,360 --> 00:17:33,760 SO WHY IS THIS IMPORTANT? 354 00:17:33,760 --> 00:17:37,360 WE HAVE THIS BIZARRE 355 00:17:37,360 --> 00:17:42,120 RELATIONSHIP RPA, RAD 52 MRE11 356 00:17:42,120 --> 00:17:42,880 IS IMPORTANT. 357 00:17:42,880 --> 00:17:44,480 WHERE IS IT NEEDED. 358 00:17:44,480 --> 00:17:46,760 AND WE SPENT LOTS OF TIME 359 00:17:46,760 --> 00:17:48,040 TRYING TO FIGURE OUT. 360 00:17:48,040 --> 00:17:50,640 A FEATURE THAT IS WORSENED BY 361 00:17:50,640 --> 00:17:52,480 DECREASING POL THETA, NOT 362 00:17:52,480 --> 00:17:54,360 WORSENED BY LOW CONCENTRATION 363 00:17:54,360 --> 00:17:57,640 OF RAD 52 INHIBITION BUT 364 00:17:57,640 --> 00:17:58,600 RESCUED WHEN WE COMBINED THE 365 00:17:58,600 --> 00:17:59,280 TWO. 366 00:17:59,280 --> 00:18:02,520 WE LOOKED AT SEVERAL FEATURES 367 00:18:02,520 --> 00:18:03,920 REPORTED IN LITERATURE 368 00:18:03,920 --> 00:18:06,000 IDENTIFIED TO BE IMPORTANT IN 369 00:18:06,000 --> 00:18:10,280 POL THETA ACTIVITY ORAD 52. 370 00:18:10,280 --> 00:18:12,640 THE SHORT STORY IS NONE OF 371 00:18:12,640 --> 00:18:13,800 THESE FEATURES REALLY 372 00:18:13,800 --> 00:18:16,640 CORRELATED WITH OUR CRITERIA. 373 00:18:16,640 --> 00:18:20,320 NOT MADE WORSE BY POL THETA 374 00:18:20,320 --> 00:18:23,800 LOSS OR NOT RESCUED BY THE 375 00:18:23,800 --> 00:18:24,120 COMBINATION. 376 00:18:24,120 --> 00:18:25,760 IT JUST DIDN'T CORRELATE WITH 377 00:18:25,760 --> 00:18:27,160 OUR OBSERVATIONS. 378 00:18:27,160 --> 00:18:28,800 WHICH LED US TO THINK ABOUT 379 00:18:28,800 --> 00:18:32,800 SOMETHING QUITE DIFFERENT. 380 00:18:32,800 --> 00:18:38,000 THIS COMES FROM LESSANDRO, 381 00:18:38,000 --> 00:18:40,080 SUGGESTED THERE ARE PHASES 382 00:18:40,080 --> 00:18:43,920 SUBSEQUENT TO NASCENT DNA 383 00:18:43,920 --> 00:18:45,360 GENERATION. 384 00:18:45,360 --> 00:18:47,560 ONE MUCH LATER IN G2. 385 00:18:47,560 --> 00:18:52,160 TO LOOK AT THE G2M REPAIR, WHAT 386 00:18:52,160 --> 00:18:54,480 HE DID ESSENTIALLY GIVE CELLS 387 00:18:54,480 --> 00:18:56,600 NUCLEOTIDE ANALOG AS THEY ARE 388 00:18:56,600 --> 00:19:01,080 GOING THROUGH S PHASE AND PUT 389 00:19:01,080 --> 00:19:02,600 THEM IN NOCODAZOLE FOR A LONG 390 00:19:02,600 --> 00:19:05,240 PERIOD OF TIME TO PUT THEM IN 391 00:19:05,240 --> 00:19:07,440 G2M AND LAST FEW HOURS OF THAT 392 00:19:07,440 --> 00:19:11,120 PROCESS GIVE THEM A SECOND 393 00:19:11,120 --> 00:19:11,320 LABEL. 394 00:19:11,320 --> 00:19:14,280 SO ANY SYNTHESIS OF THE DNA 395 00:19:14,280 --> 00:19:16,080 THAT WAS GENERATED IN S-PHASE 396 00:19:16,080 --> 00:19:17,280 COULD BE SEEN. 397 00:19:17,280 --> 00:19:19,080 YOU GET THESE LITTLE DOTS 398 00:19:19,080 --> 00:19:20,560 ARISING. 399 00:19:20,560 --> 00:19:24,920 THE DENSITY OF THOSE PER 400 00:19:24,920 --> 00:19:27,240 KILOBASE OF LABEL TELLS YOU HOW 401 00:19:27,240 --> 00:19:28,960 MUCH YOU HAVE GOT LATER IN THE 402 00:19:28,960 --> 00:19:31,200 CELSI KELL. 403 00:19:31,200 --> 00:19:32,400 CELL CYCLE. 404 00:19:32,400 --> 00:19:34,720 WHAT WE FOUND IS CELLS LACKING 405 00:19:34,720 --> 00:19:37,080 BRCA 1 HAVE A DEFECT IN THE 406 00:19:37,080 --> 00:19:40,240 ABILITY TO PUT THE FILL-IN 407 00:19:40,240 --> 00:19:41,680 SOMETHING HE REPORTED. 408 00:19:41,680 --> 00:19:44,440 HOWEVER THIS GETS WORSE WHEN WE 409 00:19:44,440 --> 00:19:46,800 DEPLETE POL THETA AND 410 00:19:46,800 --> 00:19:48,640 INTRIGUINGLY GETS BETTER IF WE 411 00:19:48,640 --> 00:19:52,160 ALSO ADD LOW LEVEL RAD 52 412 00:19:52,160 --> 00:19:53,280 INHIBITION. 413 00:19:53,280 --> 00:19:55,120 SUGGESTING THIS IS THE POINT 414 00:19:55,120 --> 00:19:58,640 WHICH THE RAD 52 IS CAUSING THE 415 00:19:58,640 --> 00:20:03,200 ISSUE LATENT CELL CYCLE. 416 00:20:03,200 --> 00:20:05,640 SO THAT DOES CORRELATE WITH OUR 417 00:20:05,640 --> 00:20:13,360 SURVIVAL ASSAYS. 418 00:20:13,360 --> 00:20:15,880 THE NOTION WE ARE COMING TO IS 419 00:20:15,880 --> 00:20:19,360 POL THETA IS INHIBITING RAD 52 420 00:20:19,360 --> 00:20:22,560 AND THAT IS DEPLETING 421 00:20:22,560 --> 00:20:23,720 POLYMERASES LATER IN THE CELL 422 00:20:23,720 --> 00:20:31,320 CYCLE. 423 00:20:31,320 --> 00:20:34,560 WHICH IS THE -- POLYMERASES. 424 00:20:34,560 --> 00:20:35,960 THE QUESTION IS WHERE IS THE 425 00:20:35,960 --> 00:20:38,040 NEED FOR POL THETA RESCUE? 426 00:20:38,040 --> 00:20:42,000 WHAT WE SEE IS THAT IN ADDITION 427 00:20:42,000 --> 00:20:44,800 TO THESE G2N FILL-INOSINE THEY 428 00:20:44,800 --> 00:20:50,160 SISES WE SEE INCREASE GAMMA RAY. 429 00:20:50,160 --> 00:20:52,280 WE SEE CHROMOSOME BREAKS WHICH 430 00:20:52,280 --> 00:20:54,480 ARE RESCUED BY THE INHIBITION 431 00:20:54,480 --> 00:20:56,680 OF RAD 52 AND WE GET HUGE 432 00:20:56,680 --> 00:21:03,080 NUMBERS OF MICRO NUCLEI RESCUED 433 00:21:03,080 --> 00:21:03,920 BY INHIB BALDWIN COUNTY 434 00:21:03,920 --> 00:21:09,040 COMMISSION INHIBITION OF RAD 435 00:21:09,040 --> 00:21:09,600 52. 436 00:21:09,600 --> 00:21:13,600 IS IT THE SAME WITH POL THETA 437 00:21:13,600 --> 00:21:14,800 INHIBITION, THEY ARE LIKELY TO 438 00:21:14,800 --> 00:21:16,360 BE USED IN THE CLINIC. 439 00:21:16,360 --> 00:21:17,800 THE ANSWER TO THIS IS YES AND 440 00:21:17,800 --> 00:21:19,760 NO. 441 00:21:19,760 --> 00:21:28,000 SO YES, IF WE JUST TAKE 53DP1 442 00:21:28,000 --> 00:21:30,360 CELLS TREAT WITH, WE SEE THE 443 00:21:30,360 --> 00:21:32,720 SAME THINGS, SENSITIVE TO 444 00:21:32,720 --> 00:21:34,800 INHIBITOR IS REDUCED, 445 00:21:34,800 --> 00:21:38,320 SUPPRESSED BY RAD 52 INHIBITORS 446 00:21:38,320 --> 00:21:42,280 OR SRNA AND SUPPRESSED BY 447 00:21:42,280 --> 00:21:43,200 LOW-DOSE MRE 11. 448 00:21:43,200 --> 00:21:49,440 THIS IS AN EXAMPLE, P31 BRCA 1 449 00:21:49,440 --> 00:21:50,960 POSITIVE. 450 00:21:50,960 --> 00:21:54,240 IF WE GIVE THEM THE INHIBITOR 451 00:21:54,240 --> 00:21:56,760 ALONE, THEY ARE VERY SENSITIVE. 452 00:21:56,760 --> 00:22:00,480 AND DECREASING VERY LOW LEVELS 453 00:22:00,480 --> 00:22:06,600 OF RPA SIRNA WILL RESTORE THE 454 00:22:06,600 --> 00:22:07,760 LEVELS OF THE CELLS. 455 00:22:07,760 --> 00:22:10,560 AS YOU MIGHT ANTICIPATE THAT 456 00:22:10,560 --> 00:22:11,960 RESCUE IS ALSO FOUND AT THE 457 00:22:11,960 --> 00:22:17,080 LEVEL OF THE DNA SYNTHESIS, SO 458 00:22:17,080 --> 00:22:19,680 WE REINHIBIT POL THETA IN THESE 459 00:22:19,680 --> 00:22:22,920 CELLS WE SEE POOR DNA SYNTHESIS 460 00:22:22,920 --> 00:22:28,000 WE CAN RESCUE BY INHIBITION OF 461 00:22:28,000 --> 00:22:32,880 RAD 52 BUT IT'S DIFFERENT IF WE 462 00:22:32,880 --> 00:22:39,080 LOOK AT BRCA 1 OR 53BP1. 463 00:22:39,080 --> 00:22:40,840 IF WE INHIBIT IT DOESN'T WORK. 464 00:22:40,840 --> 00:22:43,840 THIS IS ONE OF THE EXAMPLES, 465 00:22:43,840 --> 00:22:45,320 NOTHING IMPROVES THE SURVIVAL 466 00:22:45,320 --> 00:22:48,520 TO THE POL THETA INHIBITORS. 467 00:22:48,520 --> 00:22:49,640 SO SOMETHING VERY DIFFERENT 468 00:22:49,640 --> 00:22:52,000 GOING ON IN CELLS LACKING BRCA 469 00:22:52,000 --> 00:22:56,160 1, THE SAME THING FOR BRCA 2 470 00:22:56,160 --> 00:22:59,360 COMPARED TO CELLS LOOKING 53BP1. 471 00:22:59,360 --> 00:23:03,320 ONE OF THE BIG DIFFERENCES IN 472 00:23:03,320 --> 00:23:05,680 CELLS LIKE 53BP1 AND BRCA, IS 473 00:23:05,680 --> 00:23:10,880 THESE DNA GAPS THAT OCCUR AFTER 474 00:23:10,880 --> 00:23:13,560 TO REPRIMING WHEN ANTICIPATES 475 00:23:13,560 --> 00:23:17,480 REPLICATION DIFFICULTY. 476 00:23:17,480 --> 00:23:21,680 SO BP1 DEFICIENT CELLS WE DON'T 477 00:23:21,680 --> 00:23:28,280 SEE GAPS GIVEN TWO LABELS AND 478 00:23:28,280 --> 00:23:29,400 NUCLEASE. 479 00:23:29,400 --> 00:23:31,440 THE BRCA BP1 CELLS WE SEE 480 00:23:31,440 --> 00:23:34,760 APPEARANCE OF THESE GAPS. 481 00:23:34,760 --> 00:23:37,080 LEADING US TO WONDER IF THIS IS 482 00:23:37,080 --> 00:23:45,320 WHAT IS PERHAPS IMPORTANT. 483 00:23:45,320 --> 00:23:48,960 HOWEVER, IF WE USE THE POL 484 00:23:48,960 --> 00:23:51,120 THETA INHIBITOR IN CONTRAST TO 485 00:23:51,120 --> 00:23:53,200 WHAT'S BEEN REPORTED BY SEVERAL 486 00:23:53,200 --> 00:23:55,080 OTHER GROUPS WE DON'T SEE A 487 00:23:55,080 --> 00:23:57,160 WORSENING OF THOSE GAPS THEY 488 00:23:57,160 --> 00:23:59,480 DON'T GET BIGGER OR DIFFERENT 489 00:23:59,480 --> 00:24:00,640 OR NOTHING REALLY CHANGES 490 00:24:00,640 --> 00:24:02,800 LEADING US TO QUESTION WHETHER 491 00:24:02,800 --> 00:24:04,200 THESE GAPS ARE IMPORTANT. 492 00:24:04,200 --> 00:24:09,440 BUT I WILL COME BACK TO THAT. 493 00:24:09,440 --> 00:24:13,800 MORE OVER, COMING BACK TO THE 494 00:24:13,800 --> 00:24:17,200 DNA SYNTHESIS FILL-IN 495 00:24:17,200 --> 00:24:26,760 EXPERIMENT, CELLS LACKING 53BP1. 496 00:24:26,760 --> 00:24:29,960 CELLS ALSO LACK BRCA 1, DNA 497 00:24:29,960 --> 00:24:32,800 SYNTHESIS IS NOT IMPROVED BY 498 00:24:32,800 --> 00:24:35,800 THE INHIBITION OF RAD52. 499 00:24:35,800 --> 00:24:40,440 THIS IS ZINT. 500 00:24:40,440 --> 00:24:42,880 -- DIFFERENT. 501 00:24:42,880 --> 00:24:48,280 IN THE BRCA CELLS YOU CANNOT 502 00:24:48,280 --> 00:24:50,280 RESTORE BY INHIBITING RAD 52. 503 00:24:50,280 --> 00:24:52,320 LEADING US TO WONDER IF IT'S 504 00:24:52,320 --> 00:24:56,400 SOMETHING SPECIAL ABOUT THE 505 00:24:56,400 --> 00:24:59,160 POLYMERASE ITSELF WHEN BRCA 1 506 00:24:59,160 --> 00:25:03,880 OR 2 IS DEFICIENT. 507 00:25:03,880 --> 00:25:06,440 THESE ALSO EXPRESS BP1. 508 00:25:06,440 --> 00:25:07,840 OBVIOUS EXPERIMENT IS WHAT 509 00:25:07,840 --> 00:25:10,200 HAPPENS IF WE DEPLETE POL THETA 510 00:25:10,200 --> 00:25:13,440 AND GIVE THE POL THETA 511 00:25:13,440 --> 00:25:15,080 INHIBITOR, AND PERHAPS WHAT YOU 512 00:25:15,080 --> 00:25:16,240 MIGHT EXPECT HAPPENS. 513 00:25:16,240 --> 00:25:19,080 JUST REMIND YOU THE INHIBITOR 514 00:25:19,080 --> 00:25:20,680 REDUCES THE ABILITY TO PERFORM 515 00:25:20,680 --> 00:25:23,320 THIS GAP TO FILL IN DNA 516 00:25:23,320 --> 00:25:25,160 SYNTHESIS. 517 00:25:25,160 --> 00:25:27,960 IT DOESN'T GET BETTER WE 518 00:25:27,960 --> 00:25:29,560 COMBINE WITH INHIBITOR, IF IT 519 00:25:29,560 --> 00:25:32,480 DOESN'T GET WORSE, WE COMBINE 520 00:25:32,480 --> 00:25:34,760 WITH POL THETA DEPLETION. 521 00:25:34,760 --> 00:25:36,920 IF WE DEPLETE POL THETA, WE 522 00:25:36,920 --> 00:25:39,000 GIVE POL THETA INHIBITOR AND WE 523 00:25:39,000 --> 00:25:41,360 GIVE THEM THE RAD 35 2 524 00:25:41,360 --> 00:25:43,640 INHIBITOR, NOW WE CAN RESCUE. 525 00:25:43,640 --> 00:25:45,160 SO I EXPLAINED THAT RIGHTLY, 526 00:25:45,160 --> 00:25:47,680 LET ME DO IT ONE MORE TIME. 527 00:25:47,680 --> 00:25:50,360 POL THETA INHIBITION, WE 528 00:25:50,360 --> 00:25:51,720 RESTRICT THE ABILITY TO FILL 529 00:25:51,720 --> 00:25:57,640 IN, IF WE DEPLETE POL THETA AND 530 00:25:57,640 --> 00:25:59,720 GIVE INHIBITOR WE CAN RESCUE, 531 00:25:59,720 --> 00:26:05,720 WHICH I THINK TELLS US THE POL 532 00:26:05,720 --> 00:26:08,080 THETA DEFICIENT IS THE 533 00:26:08,080 --> 00:26:09,360 DIFFERENCE. 534 00:26:09,360 --> 00:26:13,320 WHAT IS SPECIAL ABOUT BRCA 1 OR 535 00:26:13,320 --> 00:26:14,800 2 COUNTERING INHIBITING 536 00:26:14,800 --> 00:26:16,600 POLYMERASE THAT IS DIFFERENT TO 537 00:26:16,600 --> 00:26:18,000 THE SITUATION WITH 53BP1, 538 00:26:18,000 --> 00:26:20,280 THERE'S A COUPLE WAYS TO ASK 539 00:26:20,280 --> 00:26:21,280 THIS QUESTION. 540 00:26:21,280 --> 00:26:24,960 FORTUNATELY BECAUSE WE ARE 541 00:26:24,960 --> 00:26:28,760 USING THE 61C SYSTEM POOR FO 542 00:26:28,760 --> 00:26:32,040 SI, WE CAN RESCUE BY OVER 543 00:26:32,040 --> 00:26:33,920 EXPRESSING BARD 1 IT INTERACTS 544 00:26:33,920 --> 00:26:36,240 A LITTLE BIT, NOT GREAT, BUT 545 00:26:36,240 --> 00:26:38,120 RESCUE IT BY EVER EXPRESSING 546 00:26:38,120 --> 00:26:39,000 BARD 1. 547 00:26:39,000 --> 00:26:42,240 IF WE EXPRESS BARD 1 WE CAN 548 00:26:42,240 --> 00:26:46,800 NEARLY RESTORE THE BRCA FO SI 549 00:26:46,800 --> 00:26:49,600 TO NORMAL LEVELS. 550 00:26:49,600 --> 00:26:50,720 AND INTERROGATE DIFFERENT 551 00:26:50,720 --> 00:26:51,880 ASPECTS OF BARD 1. 552 00:26:51,880 --> 00:26:53,320 I WON'T GO THROUGH ALL THE 553 00:26:53,320 --> 00:26:55,920 CONTROLS BUT THE IMPORTANT BIT 554 00:26:55,920 --> 00:26:59,280 IS THIS, MUTANT A, THIS RAD 51 555 00:26:59,280 --> 00:27:02,000 BINDING REGION OF BARD 1 FAILS 556 00:27:02,000 --> 00:27:05,960 TO SUPPRESS RAD 52 557 00:27:05,960 --> 00:27:08,080 FOSI, OVER EXPRESSION OF WILD 558 00:27:08,080 --> 00:27:10,280 TYPE BARD 1 IS CAPABLE WITH 559 00:27:10,280 --> 00:27:11,720 CELLS TREATED WITH THE 560 00:27:11,720 --> 00:27:12,480 INHIBITOR. 561 00:27:12,480 --> 00:27:15,040 ANOTHER WAY WE CAN INTERROGATE 562 00:27:15,040 --> 00:27:17,360 IT IS BY ASKING ABOUT THE 563 00:27:17,360 --> 00:27:20,400 ABILITY TO BY PASS TO GO TO RAD 564 00:27:20,400 --> 00:27:23,160 52'S ABILITY TO LOAD RAD 51. 565 00:27:23,160 --> 00:27:29,280 THIS IS A COPY OF AN 566 00:27:29,280 --> 00:27:31,520 EXPERIMENT, SHE TOOK RPA, TO 567 00:27:31,520 --> 00:27:34,280 ONE OF THE REPEATS OF BRCA 2 568 00:27:34,280 --> 00:27:38,000 AND SHE FOUND FUSING IT TO BRCA 569 00:27:38,000 --> 00:27:40,800 4 OR 3 ONE OF THE LOADING 570 00:27:40,800 --> 00:27:43,680 REPEATS WAS NECESSARY FOR BY 571 00:27:43,680 --> 00:27:43,920 PASSING. 572 00:27:43,920 --> 00:27:46,760 SO WE USED THAT TOGETHER WITH 573 00:27:46,760 --> 00:27:50,360 THE SAME, TO THE EXTREME C 574 00:27:50,360 --> 00:27:56,720 TERMINUS BRCA 2, INTERACTING 575 00:27:56,720 --> 00:28:06,240 WITH RAD 51 OLIG. 576 00:28:06,240 --> 00:28:07,600 WHEREAS THE EXTREME C TERMINUS 577 00:28:07,600 --> 00:28:11,280 IS NOT. 578 00:28:11,280 --> 00:28:14,040 AND THE BRC4 FUSION IS CAPABLE 579 00:28:14,040 --> 00:28:19,800 OF SUPPRESSING RAD 52 FOSI, THE 580 00:28:19,800 --> 00:28:21,480 POL THETA INHIBITOR. 581 00:28:21,480 --> 00:28:24,040 AS YOU WOULD EXPECT, BOTH OF 582 00:28:24,040 --> 00:28:25,440 THESE RESTORE THE ABILITY OF 583 00:28:25,440 --> 00:28:30,400 THESE CELLS TO PERFORM THIS GAP 584 00:28:30,400 --> 00:28:32,800 SYNTHESIS IN BOTH POL THETA 585 00:28:32,800 --> 00:28:35,840 DEPLETED AND IMPORTANTLY, POL 586 00:28:35,840 --> 00:28:40,720 THETA INHIBITED CELLS. 587 00:28:40,720 --> 00:28:41,720 INTERESTINGLY, IT COMPLETELY 588 00:28:41,720 --> 00:28:43,560 RESTORES THE ABILITY OF CELLS 589 00:28:43,560 --> 00:28:48,640 TO NOT HAVE THESE S1 GAPS IN 590 00:28:48,640 --> 00:28:51,440 NASCENT DNA AND AS YOU WOULD 591 00:28:51,440 --> 00:28:53,160 EXPECT RESTORES RESISTANCE TO 592 00:28:53,160 --> 00:29:00,720 THE INHIBITOR. 593 00:29:00,720 --> 00:29:02,360 SO THOROUGHLY SPEAKING WHAT WE 594 00:29:02,360 --> 00:29:09,360 HAVE ISIN' YIN TO THE YANG 595 00:29:09,360 --> 00:29:15,000 DESCRIBED BY WOLFE NAYER AND 596 00:29:15,000 --> 00:29:18,800 THE OTHER NAME ESCAPES ME. 597 00:29:18,800 --> 00:29:23,720 BRCA 1 BRCA 2 -- JOINING IN 598 00:29:23,720 --> 00:29:28,720 EXPOSING G2 PRIOR TO MITOSIS. 599 00:29:28,720 --> 00:29:32,200 WE ARE FINDING POL THETA LIMIT 600 00:29:32,200 --> 00:29:34,480 RAD 52 AND THIS IS BEHIND THE 601 00:29:34,480 --> 00:29:37,280 TOXICITY OF POL THETA LOSS. 602 00:29:37,280 --> 00:29:41,160 AND TO A DEGREE POL THETA 603 00:29:41,160 --> 00:29:42,240 INHIBITION. 604 00:29:42,240 --> 00:29:43,640 DEPENDING ON THE BACKGROUND. 605 00:29:43,640 --> 00:29:45,080 AND JUST TO GO INTO THE DETAIL 606 00:29:45,080 --> 00:29:48,680 OF WHAT I MEAN, SO WHAT WE ARE 607 00:29:48,680 --> 00:29:55,040 SEEING IS IN BRCA COMPETENT P1 608 00:29:55,040 --> 00:29:59,560 POSITIVE CELLS THERE'S NO GAPS, 609 00:29:59,560 --> 00:30:06,240 THERE'S NO G2M GAPS EITHER. 610 00:30:06,240 --> 00:30:07,640 WHEREAS IN 53BP1 CELLS THERE 611 00:30:07,640 --> 00:30:11,840 ARE NO GAPS BUT THERE ARE G2M 612 00:30:11,840 --> 00:30:13,120 GAPS. 613 00:30:13,120 --> 00:30:14,720 WE HAVE NO IDEA. 614 00:30:14,720 --> 00:30:18,000 BUT IN THE PRESENCE OF POL 615 00:30:18,000 --> 00:30:20,600 THETA, IT'S REMOVING WHAT MIGHT 616 00:30:20,600 --> 00:30:24,080 BE TOXIC RAD 52 ENGAGEMENT IN 617 00:30:24,080 --> 00:30:28,480 THOSE GAPS ALLOWING GAP FILLING 618 00:30:28,480 --> 00:30:29,120 THROUGH TRS POLYMERASES. 619 00:30:29,120 --> 00:30:32,160 IN THE ABSENCE OF BRCA 1 OR 2 620 00:30:32,160 --> 00:30:33,960 FOR MA MATTER WE ARE SEEING 621 00:30:33,960 --> 00:30:38,000 GAPS IN NASCENT DNA AND GAPS IN 622 00:30:38,000 --> 00:30:39,880 THE G2/M WHICH IN THE PRESENCE 623 00:30:39,880 --> 00:30:44,120 OF POL THETA, REMOVING RAD 52 624 00:30:44,120 --> 00:30:46,240 BUT ABSENCE OF POL THETA 625 00:30:46,240 --> 00:30:49,920 APPEARS TO BE ENCOURAGING MRE 626 00:30:49,920 --> 00:30:51,840 11 AND COUNTERACTING SOME OF 627 00:30:51,840 --> 00:30:53,200 THE POLYMERASES. 628 00:30:53,200 --> 00:30:54,120 SOMETHING DIFFERENT IS 629 00:30:54,120 --> 00:30:55,760 HAPPENING WHEN WE HAVE 630 00:30:55,760 --> 00:30:57,440 INHIBITOR TO POL THETA. 631 00:30:57,440 --> 00:30:59,960 EITHER THE PRELIMINARIES IS 632 00:30:59,960 --> 00:31:02,080 ENGAGED EARLY ON, PERHAPS TLS 633 00:31:02,080 --> 00:31:07,160 OR LATER FOR THE G2/M GAPS AND 634 00:31:07,160 --> 00:31:08,360 POLYMERASES ITSELF UNDER THESE 635 00:31:08,360 --> 00:31:10,440 CONDITIONS WHICH IS TOXIC. 636 00:31:10,440 --> 00:31:18,200 SO THE PEOPLE TO THANK ARE 637 00:31:18,200 --> 00:31:24,840 [LISTING ON SLIDE] 638 00:31:24,840 --> 00:31:26,480 THANK YOU FOR LISTENING AND 639 00:31:26,480 --> 00:31:29,280 PUTTING UP WITH THE TREMENDOUS 640 00:31:29,280 --> 00:31:30,080 TECHNICAL ISSUES. 641 00:31:30,080 --> 00:31:32,000 I WILL DO MY BEST TO ANSWER 642 00:31:32,000 --> 00:31:37,120 YOUR QUESTIONS. 643 00:31:37,120 --> 00:31:39,440 >>Chun Zhang Yang: THANK YOU 644 00:31:39,440 --> 00:31:41,320 FOR THE AMAZING TALKS. 645 00:31:41,320 --> 00:31:42,480 FOR THE PARTICIPANTS, PLEASE 646 00:31:42,480 --> 00:31:44,320 TYPE IN YOUR QUESTIONS IN THE 647 00:31:44,320 --> 00:31:48,000 CHAT BOX SO THAT WE CAN 648 00:31:48,000 --> 00:31:54,080 ELABORATE THOSE QUESTIONS. 649 00:31:54,080 --> 00:31:55,600 SO FAR I DON'T SEE ANY 650 00:31:55,600 --> 00:31:56,640 QUESTIONS. 651 00:31:56,640 --> 00:32:00,680 >>Jo Morris: JUST TO FOLLOW, I 652 00:32:00,680 --> 00:32:02,800 EXPECT GIVEN WHAT HAPPENED. 653 00:32:02,800 --> 00:32:04,200 >>Karen Usdin: -- SAID THANK 654 00:32:04,200 --> 00:32:06,760 YOU FOR THIS INTERESTING TALK, 655 00:32:06,760 --> 00:32:17,280 BUT HAS NO QUESTION, JUST YET. 656 00:32:21,760 --> 00:32:23,800 MARCELLA LATENCIA, I HOPE I'M 657 00:32:23,800 --> 00:32:27,880 PRONOUNCING, SAYS GOOD WORK, DO 658 00:32:27,880 --> 00:32:30,720 YOU PLAN TO ANALYZE SCE? 659 00:32:30,720 --> 00:32:34,720 >>Jo Morris: DEPENDS ON 660 00:32:34,720 --> 00:32:35,000 EXCHANGES. 661 00:32:35,000 --> 00:32:36,120 WE HADN'T DONE, ACTUALLY. 662 00:32:36,120 --> 00:32:38,200 NO, IT'S NOT ON OUR LIST OF 663 00:32:38,200 --> 00:32:39,400 THINGS TO DO. 664 00:32:39,400 --> 00:32:42,000 I WOULD BE INTERESTING TO KNOW 665 00:32:42,000 --> 00:32:44,320 WHY YOU THINK THAT MIGHT BE A 666 00:32:44,320 --> 00:32:46,800 GOOD IDEA. 667 00:32:46,800 --> 00:32:48,840 >>Karen Usdin: PERHAPS WHILE 668 00:32:48,840 --> 00:32:49,800 MARCELLUS IS THINKING OF THAT 669 00:32:49,800 --> 00:32:50,880 WE CAN MOVE TO THE NEXT 670 00:32:50,880 --> 00:32:53,280 QUESTION. 671 00:32:53,280 --> 00:32:55,240 CHUN, DO YOU WANT TO TAKE IT? 672 00:32:55,240 --> 00:32:56,200 MAYBE I WILL READ THE QUESTION 673 00:32:56,200 --> 00:33:00,880 UP. 674 00:33:00,880 --> 00:33:07,840 FROM KANAA, CELLS TO SIMPLE AL 675 00:33:07,840 --> 00:33:08,280 KIN NATING AGENTS? 676 00:33:08,280 --> 00:33:11,280 >>Jo Morris: WE HAVEN'T DONE 677 00:33:11,280 --> 00:33:11,480 THAT. 678 00:33:11,480 --> 00:33:12,920 I ASSUME OTHERS HAVE, I ASSUME 679 00:33:12,920 --> 00:33:14,520 THE ANSWER IS YES. 680 00:33:14,520 --> 00:33:16,560 BUT WE HAVEN'T DONE IT, BUT I 681 00:33:16,560 --> 00:33:17,960 CAN'T DIRECTLY SAY MYSELF. 682 00:33:17,960 --> 00:33:24,200 >>Karen Usdin: NICOLA, SHE 683 00:33:24,200 --> 00:33:25,480 SAYS NICE TALK, JO. 684 00:33:25,480 --> 00:33:27,240 IS THERE A PARALLEL WITH 685 00:33:27,240 --> 00:33:31,040 RESISTANCE TO POP INHIBITORS TO 686 00:33:31,040 --> 00:33:31,960 KNOCKOUT CELLS, TRAPPING 687 00:33:31,960 --> 00:33:32,880 MECHANISM? 688 00:33:32,880 --> 00:33:33,920 >>Jo Morris: I THINK THAT'S 689 00:33:33,920 --> 00:33:34,440 RIGHT. 690 00:33:34,440 --> 00:33:38,360 I THINK THERE HAS TO BE. 691 00:33:38,360 --> 00:33:39,960 THE RAD52 INHIBITION ONLY WORKS 692 00:33:39,960 --> 00:33:43,480 WHEN WE DEPLETE THE POLYMERASE 693 00:33:43,480 --> 00:33:44,920 ITSELF AND INHIBITED CONTEXT, 694 00:33:44,920 --> 00:33:46,960 YES, I THINK THERE MUST BE A 695 00:33:46,960 --> 00:33:48,000 TRAPPING MECHANISM, VERY MUCH 696 00:33:48,000 --> 00:33:49,920 SO, YES. 697 00:33:49,920 --> 00:33:51,320 AND HELLO NICOLA. 698 00:33:51,320 --> 00:33:52,800 >>Karen Usdin: NEXT QUESTION 699 00:33:52,800 --> 00:33:56,440 IS FROM MITCH, I PRESUME MITCH 700 00:33:56,440 --> 00:33:57,400 MCVEIGH. 701 00:33:57,400 --> 00:33:59,320 FASCINATING. 702 00:33:59,320 --> 00:34:01,200 APOLOGIES IF I MISSED THIS BUT 703 00:34:01,200 --> 00:34:03,000 HAVE YOU CONSIDERED -- DOES 704 00:34:03,000 --> 00:34:04,400 IT BEHAVE LIKE THE OTHER 705 00:34:04,400 --> 00:34:04,680 INHIBITOR. 706 00:34:04,680 --> 00:34:06,560 >>Jo Morris: WE HAVE TRIED IT. 707 00:34:06,560 --> 00:34:08,680 AND WE ARE STRUGGLING WITH IT. 708 00:34:08,680 --> 00:34:10,800 I DON'T KNOW IF THAT'S THE 709 00:34:10,800 --> 00:34:11,960 EXPERIENCE FOR OTHER PEOPLE, WE 710 00:34:11,960 --> 00:34:14,800 ARE A BIT CONCERNED IT'S OFF 711 00:34:14,800 --> 00:34:16,360 TARGET EFFECTS. 712 00:34:16,360 --> 00:34:22,440 I WOULDN'T WANT TO MAKE ANY 713 00:34:22,440 --> 00:34:23,720 FIRM CONCLUSIONS. 714 00:34:23,720 --> 00:34:28,960 WE ARE HOPING TO TEST THE 715 00:34:28,960 --> 00:34:30,560 REPAIR INHIBITORS AS SOON AS 716 00:34:30,560 --> 00:34:33,440 THE MTA IS SIGNED. 717 00:34:33,440 --> 00:34:35,040 >>Chun Zhang Yang: I'M GOING 718 00:34:35,040 --> 00:34:39,640 TO THE NEXT QUESTION. 719 00:34:39,640 --> 00:34:42,480 CAN YOU ELABORATE ON THE DOSE 720 00:34:42,480 --> 00:34:44,280 DEPENDENT ON THE RAD 52 KNOCK 721 00:34:44,280 --> 00:34:44,600 DOWN. 722 00:34:44,600 --> 00:34:47,440 >>Jo Morris: YEAH, HI PHILIP. 723 00:34:47,440 --> 00:34:49,520 LOVELY TO HEAR FROM YOU. 724 00:34:49,520 --> 00:34:50,680 IT'S EXTRAORDINARY, WE HAVE 725 00:34:50,680 --> 00:34:53,440 THIS BELL CURVE. 726 00:34:53,440 --> 00:34:55,840 TOO MUCH RAD52 INHIBITOR AND WE 727 00:34:55,840 --> 00:34:57,360 HAVE AN HR DEFECT IN THESE 728 00:34:57,360 --> 00:34:58,840 CELLS. 729 00:34:58,840 --> 00:35:01,560 TOO LITTLE RAD 52 INHIBITOR, 730 00:35:01,560 --> 00:35:04,320 ALL IN THE BACKGROUND OF POL 731 00:35:04,320 --> 00:35:05,560 THETA, SORRY, WE HAVE THE 732 00:35:05,560 --> 00:35:07,640 EFFECT OF POL THETA LOSS. 733 00:35:07,640 --> 00:35:09,960 THERE SEEMS TO BE A SWEET SPOT 734 00:35:09,960 --> 00:35:11,440 OF NOT TERRIBLY MUCH. 735 00:35:11,440 --> 00:35:13,240 AND IT'S DIFFICULT WITH THE 736 00:35:13,240 --> 00:35:15,120 INHIBITOR TO SAY EXACTLY WHAT 737 00:35:15,120 --> 00:35:18,840 THAT IS, BUT WE CAN START TO 738 00:35:18,840 --> 00:35:24,440 DELVE INTO IT WHEN WE USE THE 739 00:35:24,440 --> 00:35:27,000 SIRNA, THE AMOUNT WE ARE LOSING 740 00:35:27,000 --> 00:35:30,040 IS, YOU KNOW, 5-15%. 741 00:35:30,040 --> 00:35:32,560 SO AT THAT VERY LOW LEVEL, AS 742 00:35:32,560 --> 00:35:36,040 FAR AS WE KNOW WE HAVE 743 00:35:36,040 --> 00:35:37,920 FUNCTIONAL RPA AND YET WE ARE 744 00:35:37,920 --> 00:35:40,240 ABLE TO RESCUE THE SENSITIVITY 745 00:35:40,240 --> 00:35:41,280 TO POL THETA LOSS. 746 00:35:41,280 --> 00:35:42,800 I HOPE THAT HALFWAY ANSWERS 747 00:35:42,800 --> 00:35:45,160 YOUR QUESTION. 748 00:35:45,160 --> 00:35:48,400 GOOD. 749 00:35:48,400 --> 00:35:51,840 >>Karen Usdin: AND THERE'S A 750 00:35:51,840 --> 00:35:53,480 QUESTION FROM HAYA, GREAT TALK, 751 00:35:53,480 --> 00:35:57,960 IN YOUR EXPERIENCE IS POL THETA 752 00:35:57,960 --> 00:35:59,160 ACTING AS -- 753 00:35:59,160 --> 00:36:00,440 >>Jo Morris: THAT'S A LOVELY 754 00:36:00,440 --> 00:36:01,440 QUESTION. 755 00:36:01,440 --> 00:36:02,480 I DON'T KNOW. 756 00:36:02,480 --> 00:36:04,720 I DON'T KNOW. 757 00:36:04,720 --> 00:36:07,720 I THINK IT'S TLS POLYMERASE, 758 00:36:07,720 --> 00:36:09,560 THE DOUBLE STRAND BREAKS ARE 759 00:36:09,560 --> 00:36:12,160 SUBSEQUENT TO THE ACTIVITY OF 760 00:36:12,160 --> 00:36:15,080 THE POL THETA AND RAD52. 761 00:36:15,080 --> 00:36:16,840 WE KNOW WE CAN RESCUE THE 762 00:36:16,840 --> 00:36:18,920 BREAKS SO I'M GUESSING IT'S TLS 763 00:36:18,920 --> 00:36:19,840 AT THE FORK. 764 00:36:19,840 --> 00:36:23,840 AND PROBABLY I SHOULD HAVE SAID 765 00:36:23,840 --> 00:36:28,560 THE BRC4 IS THE REGION OF 766 00:36:28,560 --> 00:36:32,360 IMPLICATED STABILIZING WITH POL 767 00:36:32,360 --> 00:36:32,560 ALPHA. 768 00:36:32,560 --> 00:36:35,120 SUGGESTS IF YOU DOSE -- 769 00:36:35,120 --> 00:36:39,680 EXTRACT WITH THAT BRC4 YOU LOSE 770 00:36:39,680 --> 00:36:40,760 THE GETTING THE REST OF THE 771 00:36:40,760 --> 00:36:41,880 GAPS. 772 00:36:41,880 --> 00:36:44,360 MY GUESS IT'S STEPPING IN AS 773 00:36:44,360 --> 00:36:46,720 TLS, THAT'S BEFORE THE PROBLEMS 774 00:36:46,720 --> 00:36:48,680 ARE ARISING, BUT IT'S A GUESS. 775 00:36:48,680 --> 00:36:52,640 >>Chun Zhang Yang: I SEE A 776 00:36:52,640 --> 00:36:55,640 QUESTION FROM SARA LE -- 777 00:36:55,640 --> 00:37:00,840 I WAS CURIOUS BECAUSE OF 778 00:37:00,840 --> 00:37:05,880 CHROMATIZE HAPPENING IN HR, 779 00:37:05,880 --> 00:37:07,280 WONDERING IF 53BP1 IN -- 780 00:37:07,280 --> 00:37:08,400 CELLS WOULD HAVE CHANGED THEM 781 00:37:08,400 --> 00:37:09,360 AROUND. 782 00:37:09,360 --> 00:37:10,520 JUST OUT OF CURIOSITY. 783 00:37:10,520 --> 00:37:12,360 >>Jo Morris: CHANGE THE AMOUNT 784 00:37:12,360 --> 00:37:12,920 OF WHAT? 785 00:37:12,920 --> 00:37:16,640 >>Chun Zhang Yang: SHE DIDN'T 786 00:37:16,640 --> 00:37:18,040 SAY EITHER. 787 00:37:18,040 --> 00:37:20,160 >>Karen Usdin: I THINK RELATED 788 00:37:20,160 --> 00:37:25,400 TO THE QUESTION OF SYSTEM 789 00:37:25,400 --> 00:37:26,480 CHROMATID EXCHANGES. 790 00:37:26,480 --> 00:37:28,600 >>Jo Morris: WOULD IT INCREASE 791 00:37:28,600 --> 00:37:29,880 THE HR, YES IT DOES. 792 00:37:29,880 --> 00:37:30,840 IF THAT'S THE QUESTION YOU ARE 793 00:37:30,840 --> 00:37:32,360 ASKING. 794 00:37:32,360 --> 00:37:34,680 IN RELATION TO WILD TYPE, OR 795 00:37:34,680 --> 00:37:39,120 THE DOUBLE MUTANT HAS MORE HR. 796 00:37:39,120 --> 00:37:41,880 >>Karen Usdin: OKAY, THERE'S A 797 00:37:41,880 --> 00:37:42,800 QUESTION FROM MICHAEL. 798 00:37:42,800 --> 00:37:43,520 GREAT TALK. 799 00:37:43,520 --> 00:37:46,520 DO YOU THINK THIS ROLL OF RAD 800 00:37:46,520 --> 00:37:48,600 52 IS DISTINCT FROM SUPPOSED 801 00:37:48,600 --> 00:37:49,800 ROLE IN SINGLE STRANDED 802 00:37:49,800 --> 00:37:51,440 KNEELING? 803 00:37:51,440 --> 00:37:52,640 >>Jo Morris: YEAH, I DON'T 804 00:37:52,640 --> 00:37:57,000 KNOW. 805 00:37:57,000 --> 00:37:58,600 YEAH, THE THING IS -- NUMBER 806 00:37:58,600 --> 00:38:01,640 OF ROLES OF RAD52 JUST GROWS BY 807 00:38:01,640 --> 00:38:03,320 THE DAY, DOESN'T IT? 808 00:38:03,320 --> 00:38:05,120 WE ARE WONDERING. 809 00:38:05,120 --> 00:38:07,640 WE CAN'T DISTINGUISH IS WHETHER 810 00:38:07,640 --> 00:38:18,160 IT'S ROLES IN RAD51 LOADING AND 811 00:38:19,920 --> 00:38:22,400 HOMOLOGOUS MIGHT ALSO RELATE, 812 00:38:22,400 --> 00:38:24,240 WE ARE PLANNING TO DRILL INTO 813 00:38:24,240 --> 00:38:25,560 THAT QUESTION, I THINK THE A 814 00:38:25,560 --> 00:38:26,400 TERRIFIC QUESTION. 815 00:38:26,400 --> 00:38:27,360 WE DON'T KNOW. 816 00:38:27,360 --> 00:38:30,120 IT ALSO WRAPS UP SINGLE 817 00:38:30,120 --> 00:38:32,720 STRANDED DNA IN A 818 00:38:32,720 --> 00:38:37,400 CONCENTRATION-DEPENDENT WAY. 819 00:38:37,400 --> 00:38:40,880 LOVELY IN VITRO WORK SUGGESTS 820 00:38:40,880 --> 00:38:44,840 TOO MUCH RAD 52 HOLD UP TOO 821 00:38:44,840 --> 00:38:46,720 MUCH SINGLE STRANDED DNA IN A 822 00:38:46,720 --> 00:38:49,760 WAY THAT MIGHT BE INCOMPATIBLE 823 00:38:49,760 --> 00:38:51,640 WITH DOWNSTREAM REPAIR EVENTS. 824 00:38:51,640 --> 00:38:54,480 SORRY, I DON'T KNOW IF I'M 825 00:38:54,480 --> 00:38:55,920 EXPLAINING THAT VERY WELL, BUT 826 00:38:55,920 --> 00:38:57,160 YEAH, WE DON'T KNOW IS THE 827 00:38:57,160 --> 00:38:57,480 SHORT ANSWER. 828 00:38:57,480 --> 00:38:58,600 >>Karen Usdin: THANK YOU. 829 00:38:58,600 --> 00:39:00,440 ARE THERE ANY MORE QUESTIONS? 830 00:39:00,440 --> 00:39:02,640 RIGHT NOW THE CHAT BOX IS 831 00:39:02,640 --> 00:39:04,840 DEPLETED. 832 00:39:04,840 --> 00:39:05,800 SO LAST-MINUTE QUESTIONS FROM 833 00:39:05,800 --> 00:39:07,680 ANYBODY? 834 00:39:07,680 --> 00:39:10,880 >>I JUST HAVE A QUICK QUESTION. 835 00:39:10,880 --> 00:39:13,680 THE POL THETA INHIBITOR, HOW 836 00:39:13,680 --> 00:39:15,080 SPECIFIC IS IT? 837 00:39:15,080 --> 00:39:17,280 IT WAS ALREADY ADDRESSED IN THE 838 00:39:17,280 --> 00:39:17,560 QUESTIONS. 839 00:39:17,560 --> 00:39:19,400 BUT THE ONE YOU USED, HOW 840 00:39:19,400 --> 00:39:21,520 SPECIFIC IS IT, ACTUALLY? 841 00:39:21,520 --> 00:39:23,480 >>Jo Morris: WELL, FROM THE 842 00:39:23,480 --> 00:39:24,520 PREVIOUS PAPERS IT LOOKS VERY 843 00:39:24,520 --> 00:39:28,000 GOOD. 844 00:39:28,000 --> 00:39:30,320 I THINK WHAT'S ENCOURAGING FROM 845 00:39:30,320 --> 00:39:32,960 OUR PERSPECTIVE WE COMBINED IT 846 00:39:32,960 --> 00:39:40,720 WITH POL THETA SIRNA AND SEE 847 00:39:40,720 --> 00:39:43,040 BEAUTIFUL STASIS. 848 00:39:43,040 --> 00:39:45,400 THEY ARE NO LONGER TERRIBLY 849 00:39:45,400 --> 00:39:46,480 SPECIFIC. 850 00:39:46,480 --> 00:39:49,080 BUT IT LOOKS PRETTY GOOD. 851 00:39:49,080 --> 00:39:51,440 THERE'S A DERIVATIVE GOING INTO 852 00:39:51,440 --> 00:39:52,080 CLINICAL TRIALS, SO I HOPE IT'S 853 00:39:52,080 --> 00:39:55,200 GOOD. 854 00:39:55,200 --> 00:39:56,920 >>Karen Usdin: OKAY. 855 00:39:56,920 --> 00:40:01,040 ANY OTHER QUESTIONS? 856 00:40:01,040 --> 00:40:02,040 NOT SEEING ANY QUESTIONS IN THE 857 00:40:02,040 --> 00:40:02,720 CHAT. 858 00:40:02,720 --> 00:40:08,760 I THINK UNFORTUNATELY CHUN HAS 859 00:40:08,760 --> 00:40:10,080 DROPPED OFF THE CALL. 860 00:40:10,080 --> 00:40:12,520 WE ARE HAVING INTERNET ISSUES 861 00:40:12,520 --> 00:40:13,680 ON THE CALL ALL AROUND. 862 00:40:13,680 --> 00:40:16,440 I THINK THIS HAS BEEN GOOD 863 00:40:16,440 --> 00:40:16,720 DISCUSSION. 864 00:40:16,720 --> 00:40:18,440 IF THERE ARE NO OTHER 865 00:40:18,440 --> 00:40:20,080 QUESTIONS, I THINK JO, WE WILL 866 00:40:20,080 --> 00:40:22,560 JUST THANK YOU SO MUCH FOR A 867 00:40:22,560 --> 00:40:23,960 REALLY, REALLY INTERESTING TALK. 868 00:40:23,960 --> 00:40:26,400 LOTS OF FOOD FOR THOUGHT. 869 00:40:26,400 --> 00:40:28,720 WE WILL BE IN TOUCH. 870 00:40:28,720 --> 00:40:30,440 WE WILL SEND YOU THE LIST OF 871 00:40:30,440 --> 00:40:31,560 THE PARTICIPANTS AND QUESTIONS. 872 00:40:31,560 --> 00:40:34,480 IF YOU WANT TO TOUCH BASE WITH 873 00:40:34,480 --> 00:40:35,640 ANYBODY, YOU KNOW, THAT'S AN 874 00:40:35,640 --> 00:40:38,800 OPPORTUNITY TO DO SO. 875 00:40:38,800 --> 00:40:40,560 STEVE JACKSON, LAST MINUTE 876 00:40:40,560 --> 00:40:43,720 INPUT SAYS GREAT TALK, JO. 877 00:40:43,720 --> 00:40:46,080 CHUN IS BACK SO MAYBE HE CAN DO 878 00:40:46,080 --> 00:40:46,640 THE SIGN-OFF FOR US. 879 00:40:46,640 --> 00:40:48,960 >>Chun Zhang Yang: SORRY, JUST 880 00:40:48,960 --> 00:40:50,040 LOGGED IN. 881 00:40:50,040 --> 00:40:50,960 ALL QUESTIONS ARE DONE? 882 00:40:50,960 --> 00:40:52,560 >>Karen Usdin: WE ARE NOT 883 00:40:52,560 --> 00:40:53,760 GETTING ANY MORE QUESTIONS. 884 00:40:53,760 --> 00:40:54,880 >>Chun Zhang Yang: OKAY, ALL 885 00:40:54,880 --> 00:40:55,640 RIGHT. 886 00:40:55,640 --> 00:40:56,800 SORRY ABOUT THAT. 887 00:40:56,800 --> 00:40:58,360 I KEPT DROPPING OFF THE WEBEX 888 00:40:58,360 --> 00:41:00,160 AS WELL. 889 00:41:00,160 --> 00:41:02,400 I GUESS THERE'S OVERWHELMING 890 00:41:02,400 --> 00:41:03,720 ISSUE. 891 00:41:03,720 --> 00:41:04,680 REGARDLESS, THANK YOU, DR. 892 00:41:04,680 --> 00:41:07,000 MORRIS FOR THE FANTASTIC TALK. 893 00:41:07,000 --> 00:41:09,080 WE REALLY APPRECIATE YOU TO 894 00:41:09,080 --> 00:41:10,960 PARTICIPATING AND GIVING US 895 00:41:10,960 --> 00:41:15,160 SUCH AMAZING TALKS. 896 00:41:15,160 --> 00:41:17,480 SO WE WILL KEEP IN CONTACT AND 897 00:41:17,480 --> 00:41:21,800 GET BACK FOR ANOTHER INTEREST 898 00:41:21,800 --> 00:41:22,560 GROUP, CONFERENCE. 899 00:41:22,560 --> 00:41:24,000 THANK YOU SO MUCH, EVERYBODY. 900 00:41:24,000 --> 00:41:24,720 >>Jo Morris: THANK YOU. 901 00:41:24,720 --> 00:41:27,000 THANK YOU FOR THE QUESTIONS. 902 00:41:27,000 --> 00:41:27,440 TAKE CARE, ALL. 903 00:41:27,440 --> 00:41:28,640 >>Chun Zhang Yang: THANK YOU. 904 00:41:28,640 --> 00:41:29,800 >>Karen Usdin: TAKE CARE, 905 00:41:29,800 --> 00:00:00,000 EVERYONE.