Development and Application of Innovative High Throughput CRISPR Screening Approaches
题 目：Development and Application of Innovative High Throughput CRISPR Screening Approaches
报告人：Dongxin Zhao, Ph.D.
Senior Scientist, Cancer Systems Biology, Pfizer
联系人：北京大学分子医学研究所 赵扬 (Tel. 6275-7123)
Our ability to unravel complex biology processes will entail a deeper understanding of the underlying driver gene regulatory networks and the temporal and spatial contexts in which they operate. Recent advances in loss-of-function screening techniques have become particularly powerful for identifying individual members of pathways. However, these single-gene screens are often unable to provide information on the interactions between pathways and the co-dependencies of genes. To overcome this challenge, we have developed a new CRISPR-Cas9 based high throughput genetic screening approach that enable combinatorial deciphering of the role of both single and interacting genes via a single high throughput experiment. Applying this technique in cancer across multiple cell lines and genetic backgrounds, we reconstructed the cell-type specific genetic networks and identified numerous therapeutically relevant synthetic lethal gene pairs. In addition, we utilized the same technology to elucidate the underlying carbohydrate metabolism network topology. We established an integrated framework, combining genetic, transcriptomic, and flux measurements, and revealed key nodes controlling redox homeostasis along the KEAP1-NRF2 signaling axis. Taken together, our foundational study of genetic interactions may provide insights in understanding of how networks of genes influence in complex biology processes, and also aid in the development of precision therapeutics via new druggable synthetic lethal interactions.