Details

Autor(en) / Beteiligte

• Aguirre, Andrew J
• Meyers, Robin M
• Weir, Barbara A
• Vazquez, Francisca
• Zhang, Cheng-Zhong
• Ben-David, Uri
• Cook, April
• Ha, Gavin
• Harrington, William F
• Doshi, Mihir B
• Kost-Alimova, Maria
• Gill, Stanley
• Xu, Han
• Ali, Levi D
• Jiang, Guozhi
• Pantel, Sasha
• Lee, Yenarae
• Goodale, Amy
• Cherniack, Andrew D
• Oh, Coyin
• Kryukov, Gregory
• Cowley, Glenn S
• Garraway, Levi A
• Stegmaier, Kimberly
• Roberts, Charles W
• Golub, Todd R
• Meyerson, Matthew
• Root, David E
• Tsherniak, Aviad
• Hahn, William C

Titel

Genomic Copy Number Dictates a Gene-Independent Cell Response to CRISPR/Cas9 Targeting

Ist Teil von

• Cancer discovery, 2016-08, Vol.6 (8), p.914-929

Ort / Verlag

United States

Erscheinungsjahr

2016

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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 G2 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. We found that the number of CRISPR/Cas9-induced DNA breaks dictates a gene-independent 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. ©2016 AACR.See related commentary by Sheel and Xue, p. 824See related article by Munoz et al., p. 900This article is highlighted in the In This Issue feature, p. 803.