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Genome Med. 2019 Apr 16;11(1):21. doi: 10.1186/s13073-019-0627-9.

CRISPR-SONIC: targeted somatic oncogene knock-in enables rapid in vivo cancer modeling.

Author information

1
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, 01605, USA.
2
The Wellcome Trust/Cancer Research UK Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK.
3
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, 01605, USA. Wen.Xue@umassmed.edu.
4
Program in Molecular Medicine, Department of Molecular, Cell and Cancer Biology, and Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, 368 Plantation Street, Worcester, MA, 01605, USA. Wen.Xue@umassmed.edu.

Abstract

CRISPR/Cas9 has revolutionized cancer mouse models. Although loss-of-function genetics by CRISPR/Cas9 is well-established, generating gain-of-function alleles in somatic cancer models is still challenging because of the low efficiency of gene knock-in. Here we developed CRISPR-based Somatic Oncogene kNock-In for Cancer Modeling (CRISPR-SONIC), a method for rapid in vivo cancer modeling using homology-independent repair to integrate oncogenes at a targeted genomic locus. Using a dual guide RNA strategy, we integrated a plasmid donor in the 3'-UTR of mouse β-actin, allowing co-expression of reporter genes or oncogenes from the β-actin promoter. We showed that knock-in of oncogenic Ras and loss of p53 efficiently induced intrahepatic cholangiocarcinoma in mice. Further, our strategy can generate bioluminescent liver cancer to facilitate tumor imaging. This method simplifies in vivo gain-of-function genetics by facilitating targeted integration of oncogenes.

KEYWORDS:

CRISPR; Liver cancer; Mouse model; Oncogene; RAS

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