CRISPR-mediated direct mutation of cancer genes in the mouse liver
Open accessAbstract
The study of cancer genes in mouse models has traditionally relied on genetically-engineered strains made via transgenesis or gene targeting in embryonic stem cells. Here we describe a new method of cancer model generation using the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins) system in vivo in wild-type mice. We used hydrodynamic injection to deliver a CRISPR plasmid DNA expressing Cas9 and single guide RNAs (sgRNAs) to the liver that directly target the tumour suppressor genes Pten (ref. 5) and p53 (also known as TP53 and Trp53) (ref. 6), alone and in combination. CRISPR-mediated Pten mutation led to elevated Akt phosphorylation and lipid accumulation in hepatocytes, phenocopying the effects of deletion of the gene using Cre-LoxP technology. Simultaneous targeting of Pten and p53 induced liver tumours that mimicked those caused by Cre-loxP-mediated deletion of Pten and p53. DNA sequencing of liver and tumour tissue revealed insertion or deletion mutations of the tumour suppressor genes, including bi-allelic mutations of both Pten and p53 in tumours. Furthermore, co-injection of Cas9 plasmids harbouring sgRNAs targeting the β-catenin gene and a single-stranded DNA oligonucleotide donor carrying activating point mutations led to the generation of hepatocytes with nuclear localization of β-catenin. This study demonstrates the feasibility of direct mutation of tumour suppressor genes and oncogenes in the liver using the CRISPR/Cas system, which presents a new avenue for rapid development of liver cancer models and functional genomics.
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Cdk1 or Pten were used as controls with predictable influences on cancer.
“Cdk1 or Pten were used as controls with predictable influences on cancer (36, 37).”
From The origin of hepatocellular carcinoma depends on metabolic zonation (2026) · cites this paper as [37]
The paper’s study description (section labeled Abstract; no page given) identifies Pten as a tumor-suppressor target and reports that its CRISPR-mediated mutation produced effects resembling known Pten deletion. It does not describe Pten as a control: sgGFP served as the control, while Pten-knockout mice provided a comparison. The cited paper does not address Cdk1. (LLM, conf=0.85)
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- Authors
- Xue, W., Chen, S., Yin, H., Tammela, T., Papagiannakopoulos, T., Joshi, N. S., Cai, W., Yang, G., Bronson, R., Crowley, D. G., Zhang, F., Anderson, D. G., Sharp, P. A., & Jacks, T.
- Affiliations
- Koch Institute for Integrative Cancer Research At MIT (US)
- Tufts University (US)
- Broad Institute (US)
- Howard Hughes Medical Institute (US)
- Corresponding author
- Jacks, Tyler
- ORCID
- Venue
- Nature
- Published
- 2014-08-06
- Volume
- 514
- Issue
- 7522
- Pages
- 380-384
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC4199937
- OpenAlex
- W2119771879
- Cited by
- 776
- References
- 12
- Citations by year
- 2026: 19 · 2025: 40 · 2024: 47 · 2023: 65 · 2022: 62 · 2021: 71 · 2020: 63 · 2019: 77 · 2018: 63 · 2017: 88 · 2016: 85 · 2015: 80 · 2014: 15 · 2013: 1
- FWCI
- 36.65
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
- RePEc: Research Papers in Economics (submittedVersion)
- DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc)
- PMC
- Funders
- Howard Hughes Medical Institute
- National Institutes of Health (4r37eb000244-34, 1r01ca132091-01)
- National Institute of Mental Health (DP1MH100706)
- National Cancer Institute (K99CA169512, 5‑U54‑CA151884‑04, R01CA115527, 2-P01-CA42063, R01CA133404, U54CA151884, P01CA042063, R00CA169512, R01-CA132091, P30CA014051, P30-CA-14051)
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK097768)
- National Institute of Biomedical Imaging and Bioengineering (R01EB000244, R37-EB000244)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer Genomics and Diagnostics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- CRISPR/Cas9, in vivo gene editing, hydrodynamic injection, liver cancer model, PTEN, p53, tumor suppressor, oncogenes, β-catenin, Akt phosphorylation, lipid accumulation, hepatocytes, functional genomics, single guide RNA
- MeSH terms
- Animals, Base Sequence, Cell Transformation, Neoplastic, Female, Genetic Engineering, Liver, Liver Neoplasms, Molecular Sequence Data, Mutation, Oncogenes, Phosphorylation, Genes, Tumor Suppressor, Genes, p53, Mutagenesis, Hepatocytes, Lipid Metabolism, Proto-Oncogene Proteins c-akt, PTEN Phosphohydrolase, beta Catenin, Mice, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems
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