1 00:00:05,920 --> 00:00:07,480 >> GOOD AFTERNOON, LADIES AND 2 00:00:07,480 --> 00:00:08,880 GENTLEMEN. 3 00:00:08,880 --> 00:00:10,920 TODAY I'LL TALK ONE OF MY 4 00:00:10,920 --> 00:00:17,600 RESEARCH PROJECTS INCLUDING THE 5 00:00:17,600 --> 00:00:20,000 CSB AND ACCELERATING CELLULAR 6 00:00:20,000 --> 00:00:23,680 AGING. 7 00:00:23,680 --> 00:00:24,360 EVERYONE GETS OLD. 8 00:00:24,360 --> 00:00:27,840 WE DON'T WANT TO BUT IT'S 9 00:00:27,840 --> 00:00:29,920 INEVITABLE UNLESS YOU CAN STOP 10 00:00:29,920 --> 00:00:30,120 TIME. 11 00:00:30,120 --> 00:00:31,640 SO SCIENTISTS WORK REALLY 12 00:00:31,640 --> 00:00:33,800 HARD -- 13 00:00:33,800 --> 00:01:49,720 [NO AUDIO] EPIGENETICS ARE A RESEARCH 14 00:01:49,720 --> 00:01:50,120 FIELD THAT STUDY 15 00:01:50,120 --> 00:02:02,880 [NO AUDIO] 16 00:02:02,880 --> 00:02:06,440 >> TO FORM NUCLEAR SCAFFOLD TO 17 00:02:06,440 --> 00:02:08,800 GET INTO THE NUCLEAR. 18 00:02:08,800 --> 00:02:12,920 SO IT IS WELL KNOWN THESE HAVE 19 00:02:12,920 --> 00:02:17,960 SPECIFIC STRUCTURES WHICH 20 00:02:17,960 --> 00:02:21,600 PROTRUDE INTO NUCLEARPLASM AND 21 00:02:21,600 --> 00:02:23,280 PRONE TO VARIOUS TRANSLATIONAL 22 00:02:23,280 --> 00:02:28,160 MODE FICTION SUCH AS 23 00:02:28,160 --> 00:02:29,520 PHOSPHORYLATIONS AND ONE OF THE 24 00:02:29,520 --> 00:02:31,240 OUTCOMES OF THE MODIFICATIONS IS 25 00:02:31,240 --> 00:02:34,920 THAT IT ALTER THE ELECTROSTATIC 26 00:02:34,920 --> 00:02:38,520 NATURE OF THE PROTEINS THERE BE 27 00:02:38,520 --> 00:02:39,920 REGULATING THE INTERACTION FORCE 28 00:02:39,920 --> 00:02:43,240 OF DNA MOLECULES. 29 00:02:43,240 --> 00:02:46,320 BY THAT IT BECOMES LOOSE AND 30 00:02:46,320 --> 00:02:48,240 CONDENSE AND REGULATES THE 31 00:02:48,240 --> 00:02:49,920 TRANSCRIPTION STATUS OF THE GENE 32 00:02:49,920 --> 00:02:54,080 INSIDE THESE REGIONS. 33 00:02:54,080 --> 00:02:58,720 SO, WHEN THE CHROMATIN STRUCTURE 34 00:02:58,720 --> 00:03:01,600 BECOMES LOOSE THEY CAN ACCESS 35 00:03:01,600 --> 00:03:05,320 THE DNA REGIONS THERE BE 36 00:03:05,320 --> 00:03:06,960 TRANSCRIPTION IS SWITCHED ON. 37 00:03:06,960 --> 00:03:10,920 OTHERWISE IT CANNOT REACH THESE 38 00:03:10,920 --> 00:03:17,920 DNA REGIONS. 39 00:03:17,920 --> 00:03:33,720 AND THERE'S TRIMETHYLATION AND 40 00:03:33,720 --> 00:03:40,640 THESE ARE TWO CATEGORIES. 41 00:03:40,640 --> 00:03:43,200 THIS MODEL OF AGING WAS PROPOSED 42 00:03:43,200 --> 00:03:50,520 MORE THAN TO -- 20 YEARS AGO AND 43 00:03:50,520 --> 00:03:54,080 THE CHROMATIN STRUCTURE WAS 44 00:03:54,080 --> 00:03:59,920 CAUSING ALTERED TRANSCRIPTION 45 00:03:59,920 --> 00:04:02,720 PROFILE CAUSING THE AGING IN 46 00:04:02,720 --> 00:04:03,040 PHENOTYPES. 47 00:04:03,040 --> 00:04:05,000 AND SINCE THEN LOTS OF FOLLOW-UP 48 00:04:05,000 --> 00:04:08,680 STUDIES HAVE IDENTIFIED THESE 49 00:04:08,680 --> 00:04:10,440 CHROMATIN STRUCTURES GETTING 50 00:04:10,440 --> 00:04:14,120 LOOSE AND THE TRIMETHYLATION IS 51 00:04:14,120 --> 00:04:16,280 DECREASING BY SMALL LEVELS AND 52 00:04:16,280 --> 00:04:22,160 CELLULAR LEVELS. 53 00:04:22,160 --> 00:04:27,320 AND THE EXPRESSION WAS DECREASED 54 00:04:27,320 --> 00:04:29,200 BY AGING. 55 00:04:29,200 --> 00:04:32,000 IT'S BEEN INTERESTING THAT THESE 56 00:04:32,000 --> 00:04:37,560 AGING SYNDROMES HAVE ALSO 57 00:04:37,560 --> 00:04:45,120 REPORTED TO HAVING DECREASED 58 00:04:45,120 --> 00:04:45,880 HEMATOCHROMATIN STRUCTURE AS 59 00:04:45,880 --> 00:04:51,960 WELL AND WITH THE LOSS OR 60 00:04:51,960 --> 00:04:54,520 ALTERED MODIFICATION RESULTS IN 61 00:04:54,520 --> 00:04:58,880 LOSS OF TRANSCRIPTION PRECISION 62 00:04:58,880 --> 00:05:03,200 WHICH IS CAUSING INSTABILITY 63 00:05:03,200 --> 00:05:08,440 WHICH IS VERY TOXIC TO CELLS. 64 00:05:08,440 --> 00:05:12,960 IT IS VERY WELL KNOWN IN THE 65 00:05:12,960 --> 00:05:15,520 RUNNER SYNDROME CELLS THESE ARE 66 00:05:15,520 --> 00:05:19,800 IMPORTED BUT IT WASN'T THE CASE 67 00:05:19,800 --> 00:05:20,800 IN THE COKIN SYNDROME. 68 00:05:20,800 --> 00:05:23,080 SO WHAT IS THE COKIN SYNDROME? 69 00:05:23,080 --> 00:05:26,520 IT'S A RECESSSIVE DISEASE 70 00:05:26,520 --> 00:05:28,640 ACCELERATED AGING DISORDER 71 00:05:28,640 --> 00:05:32,000 CHARACTERIZED BY 72 00:05:32,000 --> 00:05:41,280 NEURODEGENERATION, TROFISM AND 73 00:05:41,280 --> 00:05:44,000 MOUSE MODELS HAVE DYSFUNCTION 74 00:05:44,000 --> 00:05:46,720 WHICH IS CHARACTERIZED BY 75 00:05:46,720 --> 00:05:51,560 INCREASED ENERGY TURNOVER OR 76 00:05:51,560 --> 00:05:53,520 PHOSPHORYLATION AND ACCUMULATION 77 00:05:53,520 --> 00:05:58,000 OF DAMAGED MITOCHONDRIA OR ATP 78 00:05:58,000 --> 00:06:10,440 CONSUMPTION. 79 00:06:10,440 --> 00:06:12,720 THESE TWO GENES HAVE DISTINCT 80 00:06:12,720 --> 00:06:15,720 STRUCTURE BUT THE MUTATION HAS 81 00:06:15,720 --> 00:06:17,160 VERY SIMILAR PHENOTYPES. 82 00:06:17,160 --> 00:06:23,240 AND THE MAJORITY OF THE SYNDROME 83 00:06:23,240 --> 00:06:33,640 IS ALSO KNOWN PATIENT SUFFERING 84 00:06:33,640 --> 00:06:36,520 MORE AND THEY INDUCE DAMAGE 85 00:06:36,520 --> 00:06:55,400 REPAIR AND SO IN 1936 THE 86 00:06:55,400 --> 00:06:57,680 COCKAYNE SYNDROME WAS DISCOVERED 87 00:06:57,680 --> 00:07:01,280 TO HAVE REGULATION AND MOST 88 00:07:01,280 --> 00:07:08,960 RECENTLY IN 2017 CSB IS DIRECTLY 89 00:07:08,960 --> 00:07:14,200 REGULATING THE TRANSCRIPTION OF 90 00:07:14,200 --> 00:07:21,560 THE TRANSCRIPTION DEPRESSOR BY 91 00:07:21,560 --> 00:07:31,240 DEGRADING THESE REPRESSORS ATF3 92 00:07:31,240 --> 00:07:31,560 REPRESSORS. 93 00:07:31,560 --> 00:07:33,360 AND THIS HAS SO MUCH 94 00:07:33,360 --> 00:07:34,640 PHOSPHORYLATION THE REASON IS 95 00:07:34,640 --> 00:07:39,920 WHEN THERE IS DAMAGE, PART ONE 96 00:07:39,920 --> 00:07:43,160 IS RECRUITING TO THE DNA DAMAGE 97 00:07:43,160 --> 00:07:46,920 AND LOOKING AT THE 98 00:07:46,920 --> 00:07:47,760 PHOSPHORYLATION BRANCHES AND 99 00:07:47,760 --> 00:07:50,760 IT'S KNOWN TO REPLACE -- 100 00:07:50,760 --> 00:07:53,600 DISPLACE THEM FROM DAMAGED 101 00:07:53,600 --> 00:07:53,880 CHROMATIN. 102 00:07:53,880 --> 00:07:56,440 BUT WHEN THERE'S NO FUNCTIONAL 103 00:07:56,440 --> 00:07:59,160 CSB SUCH AS COCKAYNE SYNDROME 104 00:07:59,160 --> 00:08:02,280 CELLS IT EXISTS IN THE DAMAGED 105 00:08:02,280 --> 00:08:10,280 DNA WHICH CONSUMES SO MUCH OF 106 00:08:10,280 --> 00:08:12,800 CELLULAR CHANGE ASSOCIATED WITH 107 00:08:12,800 --> 00:08:13,120 PHENOTYPES. 108 00:08:13,120 --> 00:08:16,320 AND DESPITE THE IMPORTANCE THE 109 00:08:16,320 --> 00:08:18,200 FUTURE OF THESE ARE SO UNSTABLE 110 00:08:18,200 --> 00:08:23,080 AND IT'S HARD TO DETECT BY THE 111 00:08:23,080 --> 00:08:24,840 METHOD USING COMMERCIALLY 112 00:08:24,840 --> 00:08:29,120 AVAILABLE ANTIBODIES AND HARD TO 113 00:08:29,120 --> 00:08:33,040 DO SOMETHING LIKE CHP SEQUENCING 114 00:08:33,040 --> 00:08:39,320 BUT TECHNOLOGY WAS DEVELOPED AND 115 00:08:39,320 --> 00:08:43,000 THEY HAVE IDENTIFIED OKAY DATIVE 116 00:08:43,000 --> 00:08:46,680 DAMAGE INDUCED AND THEY'RE 117 00:08:46,680 --> 00:08:50,080 MOSTLY ENRICHED IN DENSE 118 00:08:50,080 --> 00:08:51,320 CHROMATIN REGIONSES. 119 00:08:51,320 --> 00:08:53,520 SO BASED ON THIS -- REGIONS. 120 00:08:53,520 --> 00:08:55,040 SO BASED ON THIS FACT, SOME 121 00:08:55,040 --> 00:08:57,680 QUESTIONS AROSE IN MY MIND. 122 00:08:57,680 --> 00:09:01,040 IS HEMATOCHROMATIN STRUCTURE 123 00:09:01,040 --> 00:09:03,000 DESTROYED IN THE CELLS AND IF 124 00:09:03,000 --> 00:09:05,120 YES DO THEY CORRELATE WITH THE 125 00:09:05,120 --> 00:09:09,760 CELLS AND WHAT IS THE CAUSE OF 126 00:09:09,760 --> 00:09:10,800 HEMATOCHROMATIN DISRUPTION IN 127 00:09:10,800 --> 00:09:14,640 THE CELLS AND WHICH DNA HAVE THE 128 00:09:14,640 --> 00:09:15,760 MOST IN THE CELLS. 129 00:09:15,760 --> 00:09:19,040 SO I STARTED AN EXPERIMENT GREW 130 00:09:19,040 --> 00:09:23,880 THE CELLS THAT ARE CSB MUTATED 131 00:09:23,880 --> 00:09:29,000 AND THESE ARE FROM CSA TO CSB 132 00:09:29,000 --> 00:09:31,840 PATIENT FIBROBLASTS. 133 00:09:31,840 --> 00:09:35,400 AND THERE ARE TWO COMPLEMENTARY 134 00:09:35,400 --> 00:09:39,920 CONTROL CELLS COMPENSATED BY THE 135 00:09:39,920 --> 00:09:41,880 WILD TYPE COUNTERPARTS. 136 00:09:41,880 --> 00:09:44,760 I ALWAYS NOTICE THE PATIENT 137 00:09:44,760 --> 00:09:47,720 CELLS HAD ENLARGED NUCLEUS. 138 00:09:47,720 --> 00:09:52,160 YOU CAN SEE THE SIZE HAS BEEN 139 00:09:52,160 --> 00:09:55,320 INCREASED IN CSB OR CSA PATIENT 140 00:09:55,320 --> 00:09:58,120 CELLS BUT THE INTENSITY WHICH 141 00:09:58,120 --> 00:10:02,520 REPRESENTS THE DENSITY OF 142 00:10:02,520 --> 00:10:04,920 NUCLEAR DNA THE MUTATED CELLS 143 00:10:04,920 --> 00:10:08,800 HAVE SIGNIFICANTLY LESS DNA 144 00:10:08,800 --> 00:10:10,600 DENSITY COMPARED TO THE CONTROL 145 00:10:10,600 --> 00:10:11,040 CELLS. 146 00:10:11,040 --> 00:10:12,600 I SUSPECT THEY'RE UNDERGOING 147 00:10:12,600 --> 00:10:14,880 SOME KIND OF CHROMATIN 148 00:10:14,880 --> 00:10:19,000 ALTERATIONS AND SUBJECT USING 149 00:10:19,000 --> 00:10:20,720 DIFFERENT HISTOMARKS. 150 00:10:20,720 --> 00:10:26,400 YOU CAN SEE THE CSB YOU -- 151 00:10:26,400 --> 00:10:28,360 MUTATED CELLS HAVE 152 00:10:28,360 --> 00:10:30,800 TRIMETHYLATION AS A MARKER FOR 153 00:10:30,800 --> 00:10:31,080 CHROMATIN. 154 00:10:31,080 --> 00:10:34,600 AND TO TEST THE POSSIBILITY THE 155 00:10:34,600 --> 00:10:37,760 DECREASE OF TRIMETHYLATION IS A 156 00:10:37,760 --> 00:10:39,880 DIRECT CONSEQUENCE OF CSB LOST, 157 00:10:39,880 --> 00:10:42,880 I GENERATED INDUCIBLE KNOCK-DOWN 158 00:10:42,880 --> 00:10:47,960 CELLS OF CSA AND CSB INTO 159 00:10:47,960 --> 00:10:53,200 COMMONLY USED LAB SLIDES. 160 00:10:53,200 --> 00:10:55,600 YOU CAN SEE INDUCED KNOCK-DOWN 161 00:10:55,600 --> 00:11:01,360 OF CSB RESULTED IN REDUCED THE 162 00:11:01,360 --> 00:11:02,280 TRIMETHYLATION MARK IN THE 163 00:11:02,280 --> 00:11:02,560 CELLS. 164 00:11:02,560 --> 00:11:14,760 I SO -- IT LEADS TO DECOMBINING 165 00:11:14,760 --> 00:11:17,000 AND THIS IS IMPORTANT IN PATIENT 166 00:11:17,000 --> 00:11:22,840 CELLS AND A TESTED HOW THE PARTS 167 00:11:22,840 --> 00:11:25,280 ACCUMULATE ON THE CELLS BY 168 00:11:25,280 --> 00:11:29,720 REDUCING OKAY DATIVE STRESS AND 169 00:11:29,720 --> 00:11:33,920 WHEN I LOOKED AT THE CSB CELLS, 170 00:11:33,920 --> 00:11:36,920 YOU CAN SEE THAT THE CSB CELLS 171 00:11:36,920 --> 00:11:40,920 ARE REACTING MUCH MORE SENSITIVE 172 00:11:40,920 --> 00:11:43,560 COMPARED TO THE CONTROL CELLS 173 00:11:43,560 --> 00:11:46,800 UNLIKE CSA CELLS AND USING 174 00:11:46,800 --> 00:11:50,360 ANOTHER DAMAGING AGENT I 175 00:11:50,360 --> 00:11:52,040 CONFIRMED THAT THE CSB CELLS ARE 176 00:11:52,040 --> 00:11:57,880 MUCH MORE SENSITIVE TO OXIDATIVE 177 00:11:57,880 --> 00:11:58,120 ADVANTAGE. 178 00:11:58,120 --> 00:12:04,760 AND I WANTED TO SEE 179 00:12:04,760 --> 00:12:07,960 SYSTEMATICALLY USING REAL-TIME 180 00:12:07,960 --> 00:12:10,480 PCR HOW THE TRIMETHYLATION HAS 181 00:12:10,480 --> 00:12:15,920 BEEN ALTERED IN MUTATED CELLS. 182 00:12:15,920 --> 00:12:18,320 THESE THREE REPRESENTATIVE 183 00:12:18,320 --> 00:12:18,560 REGIONS. 184 00:12:18,560 --> 00:12:21,560 THIS REPRESENTS THE HOUSEKEEPING 185 00:12:21,560 --> 00:12:21,920 GENE. 186 00:12:21,920 --> 00:12:26,160 THEY ALMOST DON'T HAVE 187 00:12:26,160 --> 00:12:29,560 HETEROCHROMATIN STRUCTURE AND 188 00:12:29,560 --> 00:12:35,760 THE REGION IS A DENSE REGION. 189 00:12:35,760 --> 00:12:40,920 SO I CONFIRMED THAT THESE 190 00:12:40,920 --> 00:12:43,040 REGIONS AS WELL AS OTHER REGIONS 191 00:12:43,040 --> 00:12:50,400 IN GENERAL DNA ALL THE REGIONS 192 00:12:50,400 --> 00:12:53,960 MOST OF ALL THE REGIONS HAD 193 00:12:53,960 --> 00:12:56,280 DECREASED HETEROCHROMATIN IN THE 194 00:12:56,280 --> 00:12:56,480 CELLS. 195 00:12:56,480 --> 00:13:05,560 I TRIED TO DO THIS ASSESSMENT. 196 00:13:05,560 --> 00:13:11,680 AND WITH THE METHOD WHEN THERE 197 00:13:11,680 --> 00:13:15,840 IS DAMAGE I FIXED THEM WITH 198 00:13:15,840 --> 00:13:16,680 HIGHER PERCENTAGE COMPARED TO 199 00:13:16,680 --> 00:13:20,280 JUST CONVENTIONAL CHROMATIN AND 200 00:13:20,280 --> 00:13:28,560 USED 4% INSTEAD OF 1%. 201 00:13:28,560 --> 00:13:32,920 AND INSTEAD OF USING ANTIBODIES 202 00:13:32,920 --> 00:13:52,120 IN THE OXIDATION AND WE GAVE 203 00:13:52,120 --> 00:13:53,800 CFCR FOR SEQUENCING. 204 00:13:53,800 --> 00:13:56,120 THESE ARE THE RESULTS OF WHICH 205 00:13:56,120 --> 00:13:59,920 ARE INDUCED. 206 00:13:59,920 --> 00:14:01,680 IN THE CSB MUTATED OR CONTROLLED 207 00:14:01,680 --> 00:14:06,160 CELLS AND YOU CAN SEE THESE 208 00:14:06,160 --> 00:14:09,920 HYDROGEN PEROXIDE TREATMENT 209 00:14:09,920 --> 00:14:15,840 INDUCED PARLYLATION IN THESE 210 00:14:15,840 --> 00:14:16,080 REGIONS. 211 00:14:16,080 --> 00:14:21,280 AND I CONFIRMED THAT WITH 212 00:14:21,280 --> 00:14:25,240 ANOTHER DNA THE CSB MUTATED 213 00:14:25,240 --> 00:14:30,160 CELLS HAVE MORE ACCUMULATION BUT 214 00:14:30,160 --> 00:14:36,880 NOT IN THE CHROMATINIZED REGION. 215 00:14:36,880 --> 00:14:39,720 AND I LOOKED AT THE SEQUENCING 216 00:14:39,720 --> 00:14:51,120 TO CONFIRM THE RESULT. 217 00:14:51,120 --> 00:14:54,080 AND I COULDN'T FIND DISTRIBUTION 218 00:14:54,080 --> 00:14:55,560 BETWEEN THE CELLS WHICH IS 219 00:14:55,560 --> 00:14:57,840 DISAPPOINTING AND WHEN I DID A 220 00:14:57,840 --> 00:15:03,080 DETAILED STUDY CALCULATING PEAK 221 00:15:03,080 --> 00:15:05,480 ENRICHMENT NOT ENRICHED IN CSB 222 00:15:05,480 --> 00:15:09,880 MUTATED CELLS I COULD FIND THAT 223 00:15:09,880 --> 00:15:12,600 GSS TRANSCRIPTION FLANKING 224 00:15:12,600 --> 00:15:17,480 REGIONS ARE MORE PAR SENSITIVE 225 00:15:17,480 --> 00:15:20,320 THAN MUTATED CELLS. 226 00:15:20,320 --> 00:15:27,200 AND AS YOU CAN SEE THE CSB 227 00:15:27,200 --> 00:15:30,200 MUTATED CELLS ARE HAVING MUCH 228 00:15:30,200 --> 00:15:35,560 LESS CHROMATIN ON THE START SIDE 229 00:15:35,560 --> 00:15:37,560 THAN THE MUTATED CELLS. 230 00:15:37,560 --> 00:15:40,640 AND TO EXCLUDE THE POSSIBILITY 231 00:15:40,640 --> 00:15:44,120 THAT THESE TRANSCRIPTION SITES 232 00:15:44,120 --> 00:15:48,440 AND PEAK REGIONS, I DID THE 233 00:15:48,440 --> 00:15:50,960 CALCULATIONS AND IT TURNED OUT 234 00:15:50,960 --> 00:15:55,280 MOST OF THESE HETEROCHROMATIN 235 00:15:55,280 --> 00:15:58,920 PEAK REGIONS ARE FAR FROM 236 00:15:58,920 --> 00:15:59,840 TRANSCRIPTION START SIDE 237 00:15:59,840 --> 00:16:01,240 INCLUDING THE POSSIBILITY THESE 238 00:16:01,240 --> 00:16:06,600 TWO REGIONS ARE OVERLAPPING. 239 00:16:06,600 --> 00:16:10,160 AND I DID THE ALIGNMENT OF THE 240 00:16:10,160 --> 00:16:13,160 SEQUENCING PIECE TO THE 241 00:16:13,160 --> 00:16:15,880 TRANSCRIPTION START SITE OR THE 242 00:16:15,880 --> 00:16:19,000 REGIONS AND I GOT CONSISTENT 243 00:16:19,000 --> 00:16:23,640 RESULTS WITH REAL-TIME RESULTS. 244 00:16:23,640 --> 00:16:28,920 THE CELLS HAVE MORE PARYLATION 245 00:16:28,920 --> 00:16:33,920 AND THERE WAS MINIMAL CHANGES IN 246 00:16:33,920 --> 00:16:36,640 HETEROCHROMATIN REGIONS. 247 00:16:36,640 --> 00:16:43,160 AND AS WE TRY TO UNDERSTAND WHY 248 00:16:43,160 --> 00:16:45,440 THE CHROMATIN WAS DECREASED IN 249 00:16:45,440 --> 00:16:49,480 THESE CELLS, I SCREENED FOR THE 250 00:16:49,480 --> 00:16:52,040 FACTORS THAT AFFECT 251 00:16:52,040 --> 00:16:54,720 TRIMETHYLATION CSA AND CSB 252 00:16:54,720 --> 00:16:54,920 CELLS. 253 00:16:54,920 --> 00:17:01,880 AND I FOUND THIS THESE TWO 254 00:17:01,880 --> 00:17:04,680 METHODS WERE SIGNIFICANTLY 255 00:17:04,680 --> 00:17:09,520 DECREASED IN CSB MUTATED CELLS. 256 00:17:09,520 --> 00:17:12,360 AND WE HAD PROTEIN LEVELS THAT 257 00:17:12,360 --> 00:17:16,600 WITH DECREASED MUTATED CELLS. 258 00:17:16,600 --> 00:17:19,200 AND I TRIED TO UNDERSTAND WHY 259 00:17:19,200 --> 00:17:21,920 THESE TWO METHODS OF EXPRESSION 260 00:17:21,920 --> 00:17:25,880 WAS DECREASED IN MUTATED CELLS 261 00:17:25,880 --> 00:17:27,040 AND IN THE LITERATURE RESEARCH 262 00:17:27,040 --> 00:17:32,040 AND CAME UP WITH THE PAPER IN 263 00:17:32,040 --> 00:17:51,640 2017 AND THEY CLAIM THAT THE 264 00:17:51,640 --> 00:17:53,360 PROTEOLYSIS AND THE OCCUPANCY 265 00:17:53,360 --> 00:17:56,920 MEANS MUTATED CELLS ARE EASIER 266 00:17:56,920 --> 00:18:04,840 IN THE TARGET GENES. 267 00:18:04,840 --> 00:18:11,680 AND I CONFIRMED BY SEARCHING THE 268 00:18:11,680 --> 00:18:17,320 DATABASE AND THE TRANSCRIPTION 269 00:18:17,320 --> 00:18:18,600 FACTOR DATABASE AND FOUND IT WAS 270 00:18:18,600 --> 00:18:21,880 THE TARGET. 271 00:18:21,880 --> 00:18:29,720 AND FROM THAT DATABASE I FOUND 272 00:18:29,720 --> 00:18:33,280 THE TWO TRANSFERASES WERE BEING 273 00:18:33,280 --> 00:18:37,240 OCCUPIED BY FBF3. 274 00:18:37,240 --> 00:18:46,680 I ANALYZED THE SEQUENCING DATA 275 00:18:46,680 --> 00:18:47,960 FOUND THE BINDING TO THE 276 00:18:47,960 --> 00:18:54,400 PROMOTERS OF THE TWO GENES BUT 277 00:18:54,400 --> 00:19:02,160 NOT IN THE CONTROL CELLS. 278 00:19:02,160 --> 00:19:06,160 WE TRIED TO RECOVER THE 279 00:19:06,160 --> 00:19:07,280 HETEROCHROMATIN MARK AND SEE HOW 280 00:19:07,280 --> 00:19:10,880 IT WORKED IN DAMAGE INDUCED 281 00:19:10,880 --> 00:19:12,040 OSCILLATION ACCUMULATION. 282 00:19:12,040 --> 00:19:20,240 I GENERATED THE MUTATED CELLS 283 00:19:20,240 --> 00:19:27,600 AND WHEN I INDUCED THE TWO I CAN 284 00:19:27,600 --> 00:19:37,840 SEE IT COVERS THE 285 00:19:37,840 --> 00:19:38,200 TRIMETHYLATION. 286 00:19:38,200 --> 00:19:40,280 AND I FOUND IT IN THE INDUCED 287 00:19:40,280 --> 00:19:44,800 CELLS AND I ALSO CONFIRMED THE 288 00:19:44,800 --> 00:19:49,200 RESULTS BY REAL TIME PCR SHOWING 289 00:19:49,200 --> 00:19:52,200 THE INDUCED EXPRESSION DECREASES 290 00:19:52,200 --> 00:20:00,000 THE OSCILLATION ACCUMULATION. 291 00:20:00,000 --> 00:20:02,320 SINCE PATIENTS HAVE 292 00:20:02,320 --> 00:20:04,680 CHARACTERISTICS OF MITOCHONDRIAL 293 00:20:04,680 --> 00:20:10,120 DYSFUNCTION, I WANTED TO SEE IF 294 00:20:10,120 --> 00:20:15,800 THIS COULD RECOVER THE ABNORMAL 295 00:20:15,800 --> 00:20:25,040 MITOCHONDRIAL PHENOTYPES IN THE 296 00:20:25,040 --> 00:20:28,880 CELLS AND YOU CAN SEE IT HAS 297 00:20:28,880 --> 00:20:30,120 MUTATED CELLS AND BY THAT YOU 298 00:20:30,120 --> 00:20:37,200 CAN SEE DRAMATICALLY DECREASED 299 00:20:37,200 --> 00:20:54,320 THE OSCILLATION AND WE TEST THE 300 00:20:54,320 --> 00:20:59,560 LEVELS AND THE WHETHER WE USE 301 00:20:59,560 --> 00:21:02,880 THIS OR NOT IT HAD NO EFFECT AND 302 00:21:02,880 --> 00:21:06,520 HAS A RATE IN THE ALL THE 303 00:21:06,520 --> 00:21:10,680 PROFILES BUT WHEN YOU INDUCED 304 00:21:10,680 --> 00:21:15,920 THE SETDB1 RECEPTOR IT RECOVERS 305 00:21:15,920 --> 00:21:19,520 TO ALMOST NORMAL LEVEL WITH THE 306 00:21:19,520 --> 00:21:21,200 MITOCHONDRIAL PROFILES. 307 00:21:21,200 --> 00:21:26,000 AND FINALLY, I WANTED TO TEST IF 308 00:21:26,000 --> 00:21:31,560 THESE DECREASING PHENOTYPE IS 309 00:21:31,560 --> 00:21:34,000 ALSO TRULY IN CHRONOLOGICAL 310 00:21:34,000 --> 00:21:35,280 AGING MODEL. 311 00:21:35,280 --> 00:21:40,240 I GOT THESE FIBROBLAST CELLS 312 00:21:40,240 --> 00:21:43,360 FROM HEALTHY HUMAN CANDIDATES 313 00:21:43,360 --> 00:21:48,320 EITHER FEMALE OR MALE. 314 00:21:48,320 --> 00:21:53,920 AND STAINED IT FOR CSB OR SET 315 00:21:53,920 --> 00:21:57,280 DB1 EXPRESSION LEVEL AND THE 316 00:21:57,280 --> 00:22:04,840 LEVEL DECREASES BY AGING AND THE 317 00:22:04,840 --> 00:22:05,880 SETDB1 EXPRESSION LEVEL 318 00:22:05,880 --> 00:22:07,120 DECREASES BY AGING. 319 00:22:07,120 --> 00:22:09,120 AND NOT ONLY IN HUMAN CELLS. 320 00:22:09,120 --> 00:22:15,120 I TRIED TO TEST IF IT'S TRUE IN 321 00:22:15,120 --> 00:22:15,880 MICE. 322 00:22:15,880 --> 00:22:19,680 BRA 323 00:22:19,680 --> 00:22:21,800 BRAIN-TISSUE EXTRACTION AND THE 324 00:22:21,800 --> 00:22:26,320 HARD LEVELS ARE SIGNIFICANTLY 325 00:22:26,320 --> 00:22:28,120 INCREASING IN THIS LEVELS AND 326 00:22:28,120 --> 00:22:30,840 THE CSB LEVELS WAS DECREASED AND 327 00:22:30,840 --> 00:22:32,400 THE EXPRESSION WAS DECREASED AND 328 00:22:32,400 --> 00:22:36,680 AS EXPECTED, THE TRIMETHYLATION 329 00:22:36,680 --> 00:22:41,520 WAS DECREASED. 330 00:22:41,520 --> 00:22:46,520 SO, TO SUM UP, WHEN THERE IS 331 00:22:46,520 --> 00:22:56,840 NORMAL CSB, CSB IS SUFFICIENT 332 00:22:56,840 --> 00:23:02,160 AND THE LEVELS CAN INDUCE THE 333 00:23:02,160 --> 00:23:07,200 TRIMETHYLATION WHICH LEADS TO 334 00:23:07,200 --> 00:23:08,880 MITOCHONDRIAL HOMEOSTASIS BUT 335 00:23:08,880 --> 00:23:12,840 WHEN THERE'S NO OR LESS 336 00:23:12,840 --> 00:23:13,840 FUNCTIONAL CSB AN OLD ORGANISM 337 00:23:13,840 --> 00:23:20,360 EXPRESSION LEVELS DECREASE AND 338 00:23:20,360 --> 00:23:22,240 SETDB1 LEVEL DECREASED AND BY 339 00:23:22,240 --> 00:23:27,520 THAT THERE'S LESS TRIMETHYLATION 340 00:23:27,520 --> 00:23:30,320 LEVELS WHICH LEADS TO 341 00:23:30,320 --> 00:23:31,440 MITOCHONDRIA HOMEOSTASIS. 342 00:23:31,440 --> 00:23:33,720 NOW I CAN ANSWER THE QUESTIONS 343 00:23:33,720 --> 00:23:39,480 THEY HAD BEFORE I DID THESE 344 00:23:39,480 --> 00:23:39,760 EXPERIMENTS. 345 00:23:39,760 --> 00:23:42,120 FIRST, IT'S DISRUPTED IN IN THE 346 00:23:42,120 --> 00:23:43,240 CS CELLS. 347 00:23:43,240 --> 00:23:46,840 YES, DEFICIENT CELLS HAVE LESS 348 00:23:46,840 --> 00:23:48,720 TRIMETHYLATION LEVELS THAN 349 00:23:48,720 --> 00:23:51,000 CONTROL CELLS WHILE CSA CELLS 350 00:23:51,000 --> 00:23:53,640 ARE NOT AFFECTED. 351 00:23:53,640 --> 00:23:56,880 SO DOES IT CORRELATE WITH THE 352 00:23:56,880 --> 00:24:00,080 PATHOLOGY OF CELLS. 353 00:24:00,080 --> 00:24:02,920 I FOUND THE TRIMETHYLATION 354 00:24:02,920 --> 00:24:07,480 REDUCED THE STRESS AND INDUCED 355 00:24:07,480 --> 00:24:09,200 PARYLATION AND LOOKED AT THE 356 00:24:09,200 --> 00:24:11,720 DEFICIENT CELLS AND I FOUND THAT 357 00:24:11,720 --> 00:24:21,120 THE CSB MUTATED CELLS ARE 358 00:24:21,120 --> 00:24:27,240 EXPRESSING LESS AND BY LOOKING 359 00:24:27,240 --> 00:24:30,360 AT THE PRODUCT THE DEFICIENT 360 00:24:30,360 --> 00:24:33,360 CELLS ARE GREATLY INCREASED 361 00:24:33,360 --> 00:24:36,920 ESPECIALLY ON THE TRANSCRIPTION 362 00:24:36,920 --> 00:24:38,720 FLANKING REGIONS WHERE LEVELS 363 00:24:38,720 --> 00:24:40,440 ARE DECREASED IN THE MUTANT 364 00:24:40,440 --> 00:24:42,400 CELLS. 365 00:24:42,400 --> 00:24:45,640 AND FINALLY, I WOULD LIKE TO 366 00:24:45,640 --> 00:24:49,480 THANK DR. GORE FOR GIVING ME A 367 00:24:49,480 --> 00:24:51,120 GREAT OPPORTUNITY TO DO RESEARCH 368 00:24:51,120 --> 00:24:52,440 IN HIS WONDERFUL LAB AND GIVING 369 00:24:52,440 --> 00:24:55,720 ME THE OPPORTUNITY TO SPEAK IN 370 00:24:55,720 --> 00:24:59,680 FRONT OF THESE SCIENTISTS TODAY. 371 00:24:59,680 --> 00:25:04,280 I WOULD LIKE TO THANK OUR 372 00:25:04,280 --> 00:25:07,200 RESEARCH AND PRESENTATION AND 373 00:25:07,200 --> 00:25:12,320 WOULD LIKE TO SPECIALLY THANK 374 00:25:12,320 --> 00:25:14,240 EDWARD KIM BY STUDENT. 375 00:25:14,240 --> 00:25:15,760 HE'S SUCH A DEDICATED STUDENT 376 00:25:15,760 --> 00:25:18,720 AND I WOULD LIKE TO THANK DR. 377 00:25:18,720 --> 00:25:19,800 TYLER FOR THEIR DEDICATION IN MY 378 00:25:19,800 --> 00:25:22,240 STUDY. 379 00:25:22,240 --> 00:25:23,400 THANK YOU SO MUCH FOR YOUR 380 00:25:23,400 --> 00:25:23,680 ATTENTION. 381 00:25:23,680 --> 00:25:30,120 [APPLAUSE] 382 00:25:30,120 --> 00:25:34,880 >> ALL RIGHT. 383 00:25:34,880 --> 00:25:35,120 THANKS. 384 00:25:35,120 --> 00:25:36,320 SO DO YOU WANT TO RUN IT OR 385 00:25:36,320 --> 00:25:50,320 SHALL I DO IT? 386 00:25:50,320 --> 00:25:53,560 >> DR. LEE, WONDERFUL TALK. 387 00:25:53,560 --> 00:25:55,320 IF YOU CAN STAY THERE PEOPLE 388 00:25:55,320 --> 00:25:57,040 WILL ASK QUESTIONS FROM 389 00:25:57,040 --> 00:25:57,520 DIFFERENT SITES. 390 00:25:57,520 --> 00:25:59,520 LET'S GO TO STONY BROOK, NEW 391 00:25:59,520 --> 00:26:05,360 YORK. 392 00:26:05,360 --> 00:26:08,120 >> HI, I HAVE A QUESTION THAT'S 393 00:26:08,120 --> 00:26:11,720 KIND OF A GENERAL ONE AND MAYBE 394 00:26:11,720 --> 00:26:15,040 TOO OBVIOUS BUT IF THE CHROMATIN 395 00:26:15,040 --> 00:26:18,480 EFFECTS OF THE CSB DEFICIENCY 396 00:26:18,480 --> 00:26:20,320 ACCOUNT FOR THE ACCELERATING 397 00:26:20,320 --> 00:26:21,960 AGING PHENOTYPE WHAT IS GOING ON 398 00:26:21,960 --> 00:26:24,920 WITH CSA, WHY ARE THE PHENOTYPES 399 00:26:24,920 --> 00:26:31,960 SO CONVERGENT? 400 00:26:33,840 --> 00:26:36,240 >> SO BOTH PHENOTYPES ARE KNOWN 401 00:26:36,240 --> 00:26:43,720 TO HAVE SIMILAR PHENOTYPES BUT 402 00:26:43,720 --> 00:26:46,720 THE SYMPTOMS ARE LESS SEVERE 403 00:26:46,720 --> 00:26:48,160 THAN CSB MUTATED CELLS. 404 00:26:48,160 --> 00:26:51,520 I THINK THE CHROMATIN 405 00:26:51,520 --> 00:27:00,320 CONTRIBUTING THE DIFFERENCES IN - 406 00:27:00,320 --> 00:27:05,440 CSA INDUCED SECURITIES. 407 00:27:05,440 --> 00:27:11,280 >> COULD YOU REMIND US OF CSA 408 00:27:11,280 --> 00:27:13,240 CELLS SHOWED THE SAME 409 00:27:13,240 --> 00:27:14,760 MITOCHONDRIAL DEFECTS AS CSB? 410 00:27:14,760 --> 00:27:19,600 >> YES, THEY DO BUT I DIDN'T 411 00:27:19,600 --> 00:27:27,000 TEST IT IN MICE. 412 00:27:27,000 --> 00:27:29,720 SO ONE POSSIBILITY IS WORKING 413 00:27:29,720 --> 00:27:32,240 DIRECTLY ON MITOCHONDRIA AND ONE 414 00:27:32,240 --> 00:27:37,720 WORKS ON THE CHROMATIN EFFECTS? 415 00:27:37,720 --> 00:27:39,200 >> MAYBE. 416 00:27:39,200 --> 00:27:39,920 >> OKAY. 417 00:27:39,920 --> 00:27:42,440 VERY GOOD. 418 00:27:42,440 --> 00:27:54,920 PLEASE MUTE IN STONEY BOOK. 419 00:27:54,920 --> 00:27:56,160 LET'S GO TO PITTSBURGH. 420 00:27:56,160 --> 00:27:58,440 >> CAN YOU HEAR ME OKAY? 421 00:27:58,440 --> 00:27:59,440 >> YES. 422 00:27:59,440 --> 00:28:04,320 >> A WHILE AGO THERE WAS CSB 423 00:28:04,320 --> 00:28:06,920 INVOLVED WITH TRANSCRIPTION 424 00:28:06,920 --> 00:28:11,800 REPAIR USING A BEAUTIFUL FISH 425 00:28:11,800 --> 00:28:13,160 ASSAY AND WONDERING IF SOME OF 426 00:28:13,160 --> 00:28:18,480 THE RESULTS YOU'RE SEEING FROM 427 00:28:18,480 --> 00:28:20,480 HYDROGEN PEROXIDE CAN BE RELATED 428 00:28:20,480 --> 00:28:24,280 TO THE TRANSCRIPTION REPAIR IN 429 00:28:24,280 --> 00:28:30,120 CSB CELLS. 430 00:28:30,120 --> 00:28:34,840 >> I HAVEN'T LOOKED INTO THAT 431 00:28:34,840 --> 00:28:40,120 BUT AS YOU CAN SEE IN MY DATA, 432 00:28:40,120 --> 00:28:43,520 THE ACCUMULATION WAS MORE 433 00:28:43,520 --> 00:28:47,840 SEVERELY AT THE SITE THAN IN 434 00:28:47,840 --> 00:28:52,120 HETEROCHROMATIC REGIONS WHICH 435 00:28:52,120 --> 00:28:54,360 REPRESENTS THESE TRANSCRIPTION 436 00:28:54,360 --> 00:29:12,040 IS MORE PRONE TO CSB 437 00:29:12,040 --> 00:29:13,720 TRANSCRIPTION. 438 00:29:13,720 --> 00:29:16,680 SO I THINK IT HAS TO DO WITH 439 00:29:16,680 --> 00:29:23,520 THIS ACTIVITY. 440 00:29:23,520 --> 00:29:24,640 >> THANK YOU VERY MUCH FOR AN 441 00:29:24,640 --> 00:29:25,240 INTERESTING TALK. 442 00:29:25,240 --> 00:29:25,720 >> THANK YOU. 443 00:29:25,720 --> 00:29:27,920 >> OKAY. 444 00:29:27,920 --> 00:29:28,480 THANK YOU. 445 00:29:28,480 --> 00:29:30,320 PLEASE MUTE IN PITTSBURGH. 446 00:29:30,320 --> 00:29:33,200 LET'S GO TO NIEHS IN RESEARCH, 447 00:29:33,200 --> 00:29:40,360 TRIANGLE PARK. 448 00:29:40,360 --> 00:29:43,240 >> I HAVE A QUESTION. 449 00:29:43,240 --> 00:29:53,920 WHAT IS THE CONCENTRATION OF 450 00:29:53,920 --> 00:29:54,160 PEROXIDE. 451 00:29:54,160 --> 00:29:56,600 THE OTHER QUESTION IS ARE THERE 452 00:29:56,600 --> 00:30:02,680 ANY TRANSCRIPTIONAL MARKERS? 453 00:30:02,680 --> 00:30:07,680 >> PLEASE IDENTIFY YOURSELF AT 454 00:30:07,680 --> 00:30:07,880 NIEHS. 455 00:30:07,880 --> 00:30:18,600 >> NATALI VITORIMA. 456 00:30:18,600 --> 00:30:20,520 >> I COULDN'T HEAR YOU VERY 457 00:30:20,520 --> 00:30:20,720 CLEARLY. 458 00:30:20,720 --> 00:30:28,800 I'M NOT SURE I'M UNDERSTANDING. 459 00:30:28,800 --> 00:30:33,600 I USED THE HYDROGEN PEROXIDE FOR 460 00:30:33,600 --> 00:30:39,640 30 MINUTES. 461 00:30:39,640 --> 00:30:42,640 AND IT SEEMED IN MORE THAN 30 462 00:30:42,640 --> 00:30:44,880 MINUTES THE CELLS START TO DIE. 463 00:30:44,880 --> 00:30:49,120 I WANTED TO SEE ACUTE 464 00:30:49,120 --> 00:30:51,800 ACCUMULATION ON THE CELLS BY 465 00:30:51,800 --> 00:30:53,240 THAT CONCENTRATION. 466 00:30:53,240 --> 00:30:59,800 TO ANSWER THE SECOND QUESTION, I 467 00:30:59,800 --> 00:31:01,720 SEARCHED FOR THE CONTROVERSIES 468 00:31:01,720 --> 00:31:18,640 AND METHYLASES AND I SAW 469 00:31:18,640 --> 00:31:19,680 DISTINCT GENE EXPRESSION BUT I 470 00:31:19,680 --> 00:31:25,600 CANNOT TELL YOU AT THE MOMENT. 471 00:31:25,600 --> 00:31:27,520 >> WHAT IS THE LEVEL OF CELL 472 00:31:27,520 --> 00:31:31,720 DEATHS WHEN YOU USE THE 2.5 473 00:31:31,720 --> 00:31:34,840 PEROXIDE IN YOUR EXPERIMENTS? 474 00:31:34,840 --> 00:31:36,440 TO EXCLUDE THE POSSIBILITY OF 475 00:31:36,440 --> 00:31:51,640 SECONDARY EFFECT? 476 00:31:51,640 --> 00:31:53,080 >> THANK YOU. 477 00:31:53,080 --> 00:31:55,400 PLEASE MUTE AT NIEHS AND LET'S 478 00:31:55,400 --> 00:31:58,920 GO TO LEXINGTON, KENTUCKY. 479 00:31:58,920 --> 00:32:00,320 UNIVERSITY OF KENTUCKY, DAVID. 480 00:32:00,320 --> 00:32:02,320 >> WE'RE HERE BUT WE DON'T HAVE 481 00:32:02,320 --> 00:32:03,080 ANY QUESTIONS. 482 00:32:03,080 --> 00:32:05,080 I JUST WANT TO THANK DR. LEE FOR 483 00:32:05,080 --> 00:32:16,240 AN INTERESTING TALK. 484 00:32:16,240 --> 00:32:17,160 >> PONG LET'S GO TO 485 00:32:18,240 --> 00:32:21,000 >> OKAY, LET'S GO TO UNIVERSITY 486 00:32:21,000 --> 00:32:21,920 OF GAINESVILLE. 487 00:32:21,920 --> 00:32:23,760 >> CAN YOU COMMENT ON THE 488 00:32:23,760 --> 00:32:26,200 POSSIBILITY THAT IT'S EITHER AN 489 00:32:26,200 --> 00:32:33,000 INCREASE IN PARYLATION OR IF 490 00:32:33,000 --> 00:32:36,720 IT'S A DECREASE IN REMOVAL OF 491 00:32:36,720 --> 00:32:39,280 THE PAR AND THE OTHER QUESTION 492 00:32:39,280 --> 00:32:47,720 IS CAN YOU EXPLAIN AGAIN THE 493 00:32:47,720 --> 00:32:50,520 REDUCTION AND INCREASED PAR. 494 00:32:50,520 --> 00:32:50,640 ? 495 00:32:50,640 --> 00:33:01,160 THANK YOU. 496 00:33:01,160 --> 00:33:04,920 >> IT IS WELL KNOWN THAT FORMING 497 00:33:04,920 --> 00:33:07,920 CHROMATIN BY MINIMIZING DNA 498 00:33:07,920 --> 00:33:09,880 EXPOSURE TO THE ENVIRONMENT IS 499 00:33:09,880 --> 00:33:13,720 VERY GOOD FOR DNA PROTECTION. 500 00:33:13,720 --> 00:33:18,680 SO I THINK MORE HETEROCHROMATIN 501 00:33:18,680 --> 00:33:21,400 FORMATION IS CAUSING A MORE 502 00:33:21,400 --> 00:33:22,280 BENEFICIAL EFFECT FROM 503 00:33:22,280 --> 00:33:24,520 PROTECTING DNA FROM GETTING 504 00:33:24,520 --> 00:33:25,480 DAMAGED IN THE FIRST PLACE. 505 00:33:25,480 --> 00:33:38,120 AND WHAT'S THE SECOND QUESTION? 506 00:33:38,120 --> 00:33:42,720 >> IS IT A COUPLELATION -- 507 00:33:42,720 --> 00:33:45,360 ACCUMULATION OF PAR OR 508 00:33:45,360 --> 00:34:02,040 PARYLATION BEING DECREASED? 509 00:34:02,040 --> 00:34:03,240 >> IT MAKES MUCH MORE PAR 510 00:34:03,240 --> 00:34:11,280 ACCUMULATION. 511 00:34:11,280 --> 00:34:15,680 SO YOU STARTED TO MENTION THAT 512 00:34:15,680 --> 00:34:19,080 CSB COULD NOT LOCALIZE TO THE 513 00:34:19,080 --> 00:34:20,280 CELL SITES. 514 00:34:20,280 --> 00:34:26,720 IS THERE A SEQUENCE FOR THE 515 00:34:26,720 --> 00:34:27,000 GENE? 516 00:34:27,000 --> 00:34:50,640 CAN YOU EXPLAIN? 517 00:34:50,640 --> 00:34:53,200 >> SO WHENEVER THERE IS DNA 518 00:34:53,200 --> 00:34:56,320 DAMAGE OR REGULATED DAMAGE, CSB 519 00:34:56,320 --> 00:34:59,880 CAN BE THERE TO REPAIR THE 520 00:34:59,880 --> 00:35:03,040 DAMAGED DNA. 521 00:35:03,040 --> 00:35:06,000 BACK IN 2017 IT WAS IDENTIFIED 522 00:35:06,000 --> 00:35:09,600 CSB HAS CONSENSUS BINDING 523 00:35:09,600 --> 00:35:14,480 SEQUENCE THAT IS VERY SIMILAR TO 524 00:35:14,480 --> 00:35:17,880 INTERACTING SEQUENCE. 525 00:35:17,880 --> 00:35:27,720 >> HAVE YOU LOOK AT INHIBITERS? 526 00:35:27,720 --> 00:35:28,720 >> I HAVEN'T. 527 00:35:28,720 --> 00:35:35,640 I'M SORRY. 528 00:35:35,640 --> 00:35:37,080 >> THAT'S A QUICK ANSWER. 529 00:35:37,080 --> 00:35:39,400 WE CAN GO AROUND IF PEOPLE HAVE 530 00:35:39,400 --> 00:35:40,360 ADDITIONAL QUESTIONS. 531 00:35:40,360 --> 00:35:42,560 JUST UNMUTE AND JUMP IN. 532 00:35:42,560 --> 00:35:43,720 >> I HAVE A QUESTION FROM THE 533 00:35:43,720 --> 00:35:46,240 UNIVERSITY OF FLORIDA. 534 00:35:46,240 --> 00:35:51,800 SO YOU SHOW THAT THE EXPRESSION 535 00:35:51,800 --> 00:35:56,800 IS INTACT TO THE ATF IN THE 536 00:35:56,800 --> 00:35:59,720 CELLS AND RATHER YOU HAVE LESS 537 00:35:59,720 --> 00:36:02,480 TRIMETHYL RESIDUES OF THESE AND 538 00:36:02,480 --> 00:36:03,560 MORE DNA. 539 00:36:03,560 --> 00:36:08,000 DO YOU KNOW IF THERE'S ALSO A 540 00:36:08,000 --> 00:36:10,840 REEFFECT IN THE RECRUIT -- 541 00:36:10,840 --> 00:36:13,600 DEFECT OF THE RECRUITMENT IN THE 542 00:36:13,600 --> 00:36:20,080 CELL OR PURELY AN EXPRESSION 543 00:36:20,080 --> 00:36:20,360 DEFECT? 544 00:36:20,360 --> 00:36:26,400 >> I HAVEN'T TESTED FOR SETDB1 545 00:36:26,400 --> 00:36:32,080 RECRUITMENT TO THESE DNA BUT IN 546 00:36:32,080 --> 00:36:37,720 MY SPONSOR, THERE'S A DRE CRASE 547 00:36:37,720 --> 00:36:40,560 IN SETDB1 AND I THINK THE 548 00:36:40,560 --> 00:36:48,680 EXPRESSION LEVEL DECREASES. 549 00:36:48,680 --> 00:42:04,760 >> THANK YOU VERY MUCH.