Genome-Wide CRISPR/Cas9 Screening for Identification of Cancer Genes in Cell Lines.

Genome-Wide CRISPR/Cas9 Screening for Identification of Cancer Genes in Cell Lines. Methods Mol Biol. 2019;1907:125-136 Authors: Adelmann CH, Wang T, Sabatini DM, Lander ES Abstract In this protocol, pooled sgRNA libraries targeting thousands of genes are computationally designed, generated using microarray-based synthesis techniques, and packaged into lentiviral particles. Target cells of interest are transduced with the lentiviral sgRNA pools to generate a collection of knockout mutants-via Cas9-mediated genomic cleavage-and screened for a phenotype of interest. The relative abundance of each mutant in the population can be monitored over time through high-throughput sequencing of the integrated sgRNA expression cassettes. Using this technique, we outline strategies for the identification of cancer driver genes and genes mediating drug response. PMID: 30542996 [PubMed - in process]
Source: Mol Biol Cell - Category: Molecular Biology Authors: Tags: Methods Mol Biol Source Type: research