Multiplexed DNA tracing and RNA profiling in single cells with sequential FISH

　　报告题目： Multiplexed DNA tracing and RNA profiling in single cells with sequential FISH

　　报告人：Siyuan (Steven) Wang, Ph. D.

　　单位：Department of Chemistry and Chemical Biology, Harvard University

　　时间：2016年7月1日 上午10：00

　　地点：微生物所A203会议室

　　主持人： 黄力 研究员

　　摘要： The spatial organization of chromatin critically impacts genome function. Recent high-throughput chromosome-conformation-capture studies have revealed topologically associated domains (TADs) as a conserved feature of chromatin organization. How TADs are spatially organized in individual chromosomes remains, however, largely unknown. Here we developed an imaging method based on sequential fluorescence in situ hybridization (FISH) to directly map the spatial positions of numerous genomic regions along individual chromosomes inside the nucleus and used this method to trace the positions of TADs in four human interphase chromosomes. The method revealed non-fractal chromosome folding and polarized spatial arrangements of chromatin compartments in individual chromosomes. We have also developed a similar method called multiplexed error-robust FISH (MERFISH) to measure the abundances and localizations of numerous RNA species in single cells, and have demonstrated highly-efficient, single-molecule imaging of 1000 unique RNA species in individual cells.