Discovery of Cancer Drug Targets using RNAi Screening with Pooled Lentiviral shRNA Libraries
Conference Recording Apr 01, 2013
About the Speaker
Dr. Alex Chenchik, President/CSO of Cellecta, also a visiting scientist at the Harvard School of Public Health, started Cellecta in 2006. Alex received his Molecular Biology Ph.D. from the Institute of Molecular Biology, Moscow, Russia in 1983 and was a scientist at the National Cardiology Research Center in Moscow from 1983-1994. He has authored 44 publications and is an inventor on 10 issued patents.AbstractThis presentation will provide an overview of the recent innovations in the development of a lentiviral pooled format RNAi screening platform, in bioinformatics target prioritization, and in validation of potential drug targets in xenograft mouse models. We will present results of large-scale viability screens for cell-specific lethal and synthetic-lethal genes in a panel of blood, prostate, and breast cancer cell lines. Subsequent validation using single shRNA-expressing constructs showed that about 80% of the shRNAs identified in each complex library screen did in fact lead to cell death when transduced in cells. Analysis of the identified essential genes for known biological interactions revealed non-random clusters of interacting proteins that provide a useful strategy for prioritization of potential targets. Analysis of the lethal combinations indicates redundant, complementary, and compensatory responses in cancer cells. Genes modulating proliferation and survival in oncogenic cells have been further validated using ex vivo genetic screens with sub-pooled lentiviral shRNA libraries in xenograft mouse models. Also, we have recently developed combinatorial RNAi screening technology with bi-specific pooled shRNA libraries for the unbiased discovery and annotation of additive and synergistic synthetically lethal interactions. We will present results of combinatorial RNAi screening of DNA damage and repair genes in a breast cancer cell model.