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题目：CRISPR-based functional genomics platform in human neurons
主讲人：Ruilin Tian, Ph.D. Candidate in Dr. Martin Kampmann lab
Integrative Program in Quantitative Biology
University of California, San Francisco
时间：2019年12月26日（星期四）13:00-14:00
地点：吕志和楼B106
摘要：
While the catalog of human cell types and their gene expression profiles in normal and disease states is rapidly expanding, our understanding of human cell-type-specific gene function lags behind. Functional genomics screening is a powerful approach to systematically identify gene function in human cells. However, most of the previous genetic screens were conducted in cancer cells or immortalized cell lines. Such screening platforms have not yet been established in any differentiated, post-mitotic cell types, thus limiting potential insights into cell-type-specific roles of human genes.
As a graduate student in Martin Kampmann lab at UCSF, I developed a CRISPR-based functional genomics technology that enables first-ever large-scale genetic screens in human neurons. This platform integrates the CRISPR-interference (CRISPRi) and CRISPR-activation (CRISPRa) tools with a rapid and scalable method for human neuronal differentiation from induced pluripotent stem cells (iPSCs). This platform allows robust genetic perturbation in human neurons and can be used in genome-wide pooled screens based on cell survival or reporter levels. It can also be coupled with single-cell RNA sequencing and high-content imaging to uncover transcriptome and morphological consequences of gene knockdown in human neurons, respectively. Our results highlight the power of unbiased genetic screens in iPSC-derived differentiated cell types and provide a platform for systematic interrogation of normal and disease states of human neurons.
I will present how we developed this platform and how this platform was applied to uncover neuronal specific essential genes and genes regulating oxidative stress response in human neurons.
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