Mapping a functional cancer genome atlas of tumor suppressors in mouse liver using AAV-CRISPR–mediated direct in vivo screening
Author(s)Wang, Guangchuan; Chow, Ryan D.; Ye, Lupeng; Guzman, Christopher D.; Dai, Xiaoyun; Dong, Matthew B.; Zhang, Feng; Sharp, Phillip A; Platt, Randall Jeffrey; Chen, Sidi; ... Show more Show less
MetadataShow full item record
Cancer genomics consortia have charted the landscapes of numerous human cancers. Whereas somemutations 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 highthroughput in vivo mapping of functional variants in an autochthonous mouse model of cancer. Using adenoassociated 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 diedwithin 4 months.We usedmolecular 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.
DepartmentDavid H. Koch Institute for Integrative Cancer Research at MIT; Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Biology; Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences; McGovern Institute for Brain Research at MIT
American Association for the Advancement of Science (AAAS)
Wang, Guangchuan, Ryan D. Chow, Lupeng Ye, Christopher D. Guzman, Xiaoyun Dai, Matthew B. Dong, Feng Zhang, Phillip A. Sharp, Randall J. Platt, and Sidi Chen. “Mapping a Functional Cancer Genome Atlas of Tumor Suppressors in Mouse Liver Using AAV-CRISPR–mediated Direct in Vivo Screening.” Science Advances 4, no. 2 (February 2018): eaao5508.
Final published version