RNA Therapeutics Institute; Program in Molecular Medicine, Department of Molecular, Cell and Cancer Biology; Li Weibo Institute for Rare Diseases Research
Biochemistry, Biophysics, and Structural Biology | Bioinformatics | Cancer Biology | Cell and Developmental Biology | Genetic Phenomena | Genetics and Genomics | Investigative Techniques
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 beta-actin, allowing co-expression of reporter genes or oncogenes from the beta-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.
CRISPR, Liver cancer, Mouse model, Oncogene, RAS, UMCCTS funding
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DOI of Published Version
Genome Med. 2019 Apr 16;11(1):21. doi: 10.1186/s13073-019-0627-9. Link to article on publisher's site
Mou H, Ozata DM, Smith JL, Sheel A, Kwan S, Hough S, Kucukural A, Kennedy Z, Cao Y, Xue W. (2019). CRISPR-SONIC: targeted somatic oncogene knock-in enables rapid in vivo cancer modeling. RNA Therapeutics Institute Publications. https://doi.org/10.1186/s13073-019-0627-9. Retrieved from https://escholarship.umassmed.edu/rti_pubs/45
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This work is licensed under a Creative Commons Attribution 4.0 License.