Genome-wide CRISPR screen in a mouse model of tumor growth and metastasis

Details

• Alternative Title:
  Cell
• Personal Author:
  Chen, Sidi ; Sanjana, Neville E. ; Zheng, Kaijie ; Shalem, Ophir ; Lee, Kyungheon ; Shi, Xi ; Scott, David A. ; Song, Jun ; Pan, Jen Q. ; Weissleder, Ralph ; Lee, Hakho ; Zhang, Feng ; Sharp, Phillip A.
• Description:
  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.
• Subjects:
  Animals Carcinoma, Non-Small-Cell Lung CRISPR-Cas Systems Gene Knockout Techniques Genome-Wide Association Study Humans Lung Neoplasms Mice Neoplasm Metastasis RNA, Guide, CRISPR-Cas Systems
• Keywords:
  Cancer Evolution CRISPR/Cas9 Genome-wide Screen Metastasis
• Source:
  Cell. 2015; 160(6):1246-1260
• Pubmed ID:
  25748654
• Pubmed Central ID:
  PMC4380877
• Document Type:
  Journal Article
• Funding:
  R01-CA133404/CA/NCI NIH HHSUnited States/ ; U54 CA151884/CA/NCI NIH HHSUnited States/ ; K99 HG008171/HG/NHGRI NIH HHSUnited States/ ; 5DP1-MH100706/DP/NCCDPHP CDC HHSUnited States/ ; P01 CA042063/CA/NCI NIH HHSUnited States/ ; P30 CA014051/CA/NCI NIH HHSUnited States/ ; P30-CA14051/CA/NCI NIH HHSUnited States/ ; K99-HG008171/HG/NHGRI NIH HHSUnited States/ ; R01 CA133404/CA/NCI NIH HHSUnited States/ ; DP1 MH100706/MH/NIMH NIH HHSUnited States/ ; R01 DK097768/DK/NIDDK NIH HHSUnited States/ ; 5R01-DK097768/DK/NIDDK NIH HHSUnited States/
• Volume:
  160
• Issue:
  6
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:e520d794b2ced3fd2f60830d454bf01300f97543acd0d067322ba9b94420e027
• Download URL:
  https://stacks.cdc.gov/view/cdc/30747/cdc_30747_DS1.pdf
• File Type:
  [PDF - 2.36 MB ]