RNA Therapeutics Institute Publications

UMMS Affiliation

RNA Therapeutics Institute; Program in Molecular Medicine, Department of Molecular, Cell and Cancer Biology; Li Weibo Institute for Rare Diseases Research

Publication Date

2019-04-16

Document Type

Article

Disciplines

Biochemistry, Biophysics, and Structural Biology | Bioinformatics | Cancer Biology | Cell and Developmental Biology | Genetic Phenomena | Genetics and Genomics | Investigative Techniques

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 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.

Keywords

CRISPR, Liver cancer, Mouse model, Oncogene, RAS, UMCCTS funding

Rights and Permissions

© 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

10.1186/s13073-019-0627-9

Source

Genome Med. 2019 Apr 16;11(1):21. doi: 10.1186/s13073-019-0627-9. Link to article on publisher's site

Journal/Book/Conference Title

Genome medicine

Related Resources

Link to Article in PubMed

PubMed ID

30987660

Creative Commons License

Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

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