Abstract C161: CRISPR/Cas9 genome-wide gRNA library screening platform

• Published in: Molecular cancer therapeutics Vol. 14; no. 12_Supplement_2; p. C161
• Main Authors: Tedesco, Donato, Diehl, Paul, Makhanov, Mikhail, Baron, Sylvain, Suchkov, Dmitry, Chenchik, Alex
• Format: Journal Article
• Language: English
• Published: 01.12.2015
• DOI: 10.1158/1535-7163.TARG-15-C161
• ISSN: 1535-7163

Abstract CRISPR/Cas9 gene knock-out technology can be used as a powerful tool for large-scale functional genomic analysis in mammalian cells. With the goal to establish a cost-effective functional genomics platform for the discovery of therapeutic targets, we developed a highly functional high throughput lentiviral gRNA/CRISPR screening platform, enabling scientists to perform pooled format genome-wide CRISPR/Cas9 genetic screens. Three 55K gRNA libraries were designed to cover the whole protein-encoding human genome with a redundancy of 8 gRNAs per gene. As supporting tools, we developed protocols, reagents, and software tools for hit validation and target prioritization. Here we show the results of parallel genetic screens in a pair of isogenic CML cell lines (CML-R and CML-P), where the performance of CRISPR/Cas9 and RNAi screening platforms was compared for each cell line. Citation Format: Donato Tedesco, Paul Diehl, Mikhail Makhanov, Sylvain Baron, Dmitry Suchkov, Alex Chenchik. CRISPR/Cas9 genome-wide gRNA library screening platform. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C161.