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Sci Adv. 2018 Feb 28;4(2):eaao5508. doi: 10.1126/sciadv.aao5508. eCollection 2018 Feb.

Mapping a functional cancer genome atlas of tumor suppressors in mouse liver using AAV-CRISPR-mediated direct in vivo screening.

Wang G1,2, Chow RD1,2,3, Ye L1,2, Guzman CD1,2,4,5, Dai X1,2, Dong MB1,2,3,5, Zhang F6,7, Sharp PA8,9, Platt RJ10,11, Chen S1,2,4,12,13,14.

Author information

1
Department of Genetics, Yale University School of Medicine, New Haven, CT 06510, USA.
2
Systems Biology Institute, Yale University School of Medicine, 850 West Campus Drive, West Haven, CT 06516, USA.
3
MD-PhD Program, Yale University School of Medicine, New Haven, CT 06510, USA.
4
Immunobiology Program, Yale University School of Medicine, New Haven, CT 06510, USA.
5
Department of Immunobiology, Yale University School of Medicine, New Haven, CT 06510, USA.
6
Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard University, 75 Ames Street, Cambridge, MA 02142, USA.
7
Department of Biological Engineering, MIT, Cambridge, MA 02142, USA.
8
Koch Institute for Integrative Cancer Research, MIT, 77 Massachusetts Avenue, Cambridge, MA 02139-4307, USA.
9
Department of Biology, MIT, Cambridge, MA 02139-4307, USA.
10
Department of Biosystems Science and Engineering, ETH Zürich, Mattenstrasse 26, 4058 Basel, Switzerland.
11
Department of Chemistry, University of Basel, Petersplatz 1, 4003 Basel, Switzerland.
12
Biological and Biomedical Sciences Program, Yale University School of Medicine, West Haven, CT 06516, USA.
13
Comprehensive Cancer Center, Yale University School of Medicine, West Haven, CT 06516, USA.
14
Stem Cell Center, Yale University School of Medicine, West Haven, CT 06516, USA.

Abstract

Cancer genomics consortia have charted the landscapes of numerous human cancers. Whereas some mutations were found in classical oncogenes and tumor suppressors, others have not yet been functionally studied in vivo. To date, a comprehensive assessment of how these genes influence oncogenesis is lacking. We performed direct high-throughput in vivo mapping of functional variants in an autochthonous mouse model of cancer. Using adeno-associated viruses (AAVs) carrying a single-guide RNA (sgRNA) library targeting putative tumor suppressor genes significantly mutated in human cancers, we directly pool-mutagenized the livers of Cre-inducible CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein 9 (Cas9) mice. All mice that received the AAV-mTSG library developed liver cancer and died within 4 months. We used molecular inversion probe sequencing of the sgRNA target sites to chart the mutational landscape of these tumors, revealing the functional consequence of multiple variants in driving liver tumorigenesis in immunocompetent mice. AAV-mediated autochthonous CRISPR screens provide a powerful means for mapping a provisional functional cancer genome atlas of tumor suppressors in vivo.

PMID:
29503867
PMCID:
PMC5829971
DOI:
10.1126/sciadv.aao5508
[Indexed for MEDLINE]
Free PMC Article

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