Cell

Genetic screens are powerful tools for identifying genes responsible for diverse phenotypes. Here we describe a genome-wide CRISPR/Cas9-mediated loss-of-function screen in tumor growth and metastasis. We mutagenized a non-metastatic mouse cancer cell line using a genome-scale library with 67,405 single-guide RNAs (sgRNAs). The mutant cell pool rapidly generates metastases when transplanted into immunocompromised mice. Enriched sgRNAs in lung metastases and late-stage primary tumors were found to target a small set of genes, suggesting that specific loss-of-function mutations drive tumor growth and metastasis. Individual sgRNAs and a small pool of 624 sgRNAs targeting the top-scoring genes from the primary screen dramatically accelerate metastasis. In all of these experiments, the effect of mutations on primary tumor growth positively correlates with the development of metastases. Our study demonstrates Cas9-based screening as a robust method to systematically assay gene phenotypes in cancer evolution in vivo.5DP1-MH100706/DP/NCCDPHP CDC HHS/United States5R01-DK097768/DK/NIDDK NIH HHS/United StatesDP1 MH100706/MH/NIMH NIH HHS/United StatesK99 HG008171/HG/NHGRI NIH HHS/United StatesK99-HG008171/HG/NHGRI NIH HHS/United StatesP30 CA014051/CA/NCI NIH HHS/United StatesP30-CA14051/CA/NCI NIH HHS/United StatesR01 CA133404/CA/NCI NIH HHS/United StatesR01 DK097768/DK/NIDDK NIH HHS/United StatesR01-CA133404/CA/NCI NIH HHS/United StatesU54 CA151884/CA/NCI NIH HHS/United StatesU54 CA151884/CA/NCI NIH HHS/United States2016-03-12T00:00:00Z25748654PMC438087