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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© The Author(s). 2019 Open Access: This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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, Haiwei; Ozata, Deniz M.; Smith, Jordan L.; Sheel, Ankur; Kwan, Suet-Yan; Hough, Soren; Kucukural, Alper; Kennedy, Zachary; Cao, Yueying; and Xue, Wen, "CRISPR-SONIC: targeted somatic oncogene knock-in enables rapid in vivo cancer modeling" (2019). RNA Therapeutics Institute Publications. 45.
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This work is licensed under a Creative Commons Attribution 4.0 License.