Targeting mutant KRAS with CRISPR-Cas9 controls tumor growth
Open accessAbstract
KRAS is the most frequently mutated oncogene in human tumors, and its activating mutations represent important therapeutic targets. The combination of Cas9 and guide RNA from the CRISPR-Cas system recognizes a specific DNA sequence and makes a double-strand break, which enables editing of the relevant genes. Here, we harnessed CRISPR to specifically target mutant KRAS alleles in cancer cells. We screened guide RNAs using a reporter system and validated them in cancer cells after lentiviral delivery of Cas9 and guide RNA. The survival, proliferation, and tumorigenicity of cancer cells in vitro and the growth of tumors in vivo were determined after delivery of Cas9 and guide RNA. We identified guide RNAs that efficiently target mutant KRAS without significant alterations of the wild-type allele. Doxycycline-inducible expression of this guide RNA in KRAS-mutant cancer cells transduced with a lentiviral vector encoding Cas9 disrupted the mutant KRAS gene, leading to inhibition of cancer cell proliferation both in vitro and in vivo. Intra-tumoral injection of lentivirus and adeno-associated virus expressing Cas9 and sgRNA suppressed tumor growth in vivo, albeit incompletely, in immunodeficient mice. Expression of Cas9 and the guide RNA in cells containing wild-type KRAS did not alter cell survival or proliferation either in vitro and in vivo. Our study provides a proof-of-concept that CRISPR can be utilized to target driver mutations of cancers in vitro and in vivo.
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Allele-specific CRISPR approaches targeting mutant KRAS alleles (such as G12D or G12V) have shown promise in selectively disrupting oncogenic signaling while sparing the wild-type allele.
“In particular, allele-specific CRISPR approaches targeting mutant KRAS alleles (such as G12D or G12V) have shown promise in selectively disrupting oncogenic signaling while sparing the wild-type allele [103].”
From RAS Mutations in Advanced Colorectal Cancer: Mechanisms, Clinical Implications, and Novel Therapeutic Approaches (2025) · cites this paper as [103]
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- Authors
- Kim, W., Lee, S., Kim, H. S., Song, M., Cha, Y. H., Kim, Y.-H., Shin, J., Lee, E.-S., Joo, Y., Song, J. J., Choi, E. J., Choi, J. W., Lee, J., Kang, M., Yook, J. I., Lee, M. G., Kim, Y.-S., Paik, S., & Kim, H. (.
- Affiliations
- Yonsei University (KR)
- Seoul National University (KR)
- Chungnam National University (KR)
- ORCID
- Venue
- Genome Research
- Publisher
- Cold Spring Harbor Laboratory
- Published
- 2018-01-11
- Volume
- 28
- Issue
- 3
- Pages
- 374-382
- ISSN
- 1088-9051 (ISSN-L)
- 1549-5469
- PMC
- PMC5848616
- OpenAlex
- W2783578717
- Cited by
- 97
- References
- 32
- Citations by year
- 2026: 4 · 2025: 18 · 2024: 20 · 2023: 13 · 2022: 14 · 2021: 5 · 2020: 10 · 2019: 11 · 2018: 2
- Type
- posted content
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Genome Research (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- YUHSpace (Yonsei University Medical Library) (submittedVersion, cc-by-nc-nd)
- HighWire Press Open Archive (submittedVersion)
- Funders
- National Research Foundation of Korea (2017R1A2B3004198, 2017M3A9B4062403, 2014M3A9B6073507, 2013M3A9B4076544)
- Institute for Basic Science (IBS-R026-D1)
- Korean Health Technology R&D Project
- Ministry of Health and Welfare
- Republic of Korea (HI17C0676, HI14C1324)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Health Sciences › Medicine › Oncology › CAR-T cell therapy research
- Keywords
- CRISPR/Cas9, mutant KRAS, genome editing, lentiviral delivery, doxycycline-inducible expression, adeno-associated virus, tumor growth inhibition, tumor cell proliferation, driver mutations, immunodeficient mice, targeted genome editing
- MeSH terms
- Gene Editing, RNA, Guide, CRISPR-Cas Systems, Animals, Humans, Mutation, Neoplasms, Lentivirus, Proto-Oncogene Proteins p21(ras), Cell Line, Tumor, Cell Proliferation, Mice, CRISPR-Cas Systems
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