期刊
CANCER DISCOVERY
卷 6, 期 8, 页码 914-929出版社
AMER ASSOC CANCER RESEARCH
DOI: 10.1158/2159-8290.CD-16-0154
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资金
- Pancreatic Cancer Action Network Samuel Stroum Fellowship
- Hope Funds for Cancer Postdoctoral Fellowship
- American Society of Clinical Oncology Young Investigator Award
- Dana-Farber Cancer Institute Hale Center for Pancreatic Cancer
- Perry S. Levy Endowed Fellowship
- Harvard Catalyst and Harvard Clinical and Translational Science Center [UL1 TR001102]
- Slim Initiative for Genomic Medicine
- Carlos Slim Foundation in Mexico
- [R01 CA130988]
- [U01 CA199253]
- [U01 CA176058]
- [P01 CA154303]
- [P50 CA12700323]
The CRISPR/Cas9 system enables genome editing and somatic cell genetic screens in mammalian cells. We performed genome-scale loss-of-function screens in 33 cancer cell lines to identify genes essential for proliferation/survival and found a strong correlation between increased gene copy number and decreased cell viability after genome editing. Within regions of copy-number gain, CRISPR/Cas9 targeting of both expressed and unexpressed genes, as well as intergenic loci, led to significantly decreased cell proliferation through induction of a G(2) cell-cycle arrest. By examining single-guide RNAs that map to multiple genomic sites, we found that this cell response to CRISPR/Cas9 editing correlated strongly with the number of target loci. These observations indicate that genome targeting by CRISPR/Cas9 elicits a gene-independent antiproliferative cell response. This effect has important practical implications for the interpretation of CRISPR/Cas9 screening data and confounds the use of this technology for the identification of essential genes in amplified regions. SIGNIFICANCE: We found that the number of CRISPR/Cas9-induced DNA breaks dictates a geneindependent antiproliferative response in cells. These observations have practical implications for using CRISPR/Cas9 to interrogate cancer gene function and illustrate that cancer cells are highly sensitive to site-specific DNA damage, which may provide a path to novel therapeutic strategies. Cancer Discov; 6( 8); 914-29. (C) 2016 AACR.
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