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Unintended on-target chromosomal instability following CRISPR/Cas9 single gene targeting
Open accessAbstract
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) seminal studies in mammalian cells1–3 have resulted in gene editing being broadly adopted in basic research. However, it has become apparent that the CRISPR/Cas9 system induces unintended off- and on-target genomic alterations4 and that there is a need for stricter clone screening methods before phenotypic characterisation is made, particularly before the technology is adopted for clinical purposes.
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Przewrocka et al. observed a 6% incidence of chromosome arm truncations when targeting ZNF516 in p53-competent HCT116 cancer cell lines with CRISPR-Cas9.
“More recently, Przewrocka and colleagues observed a 6% incidence of chromosome arm truncations when targeting ZNF516 in p53-competent HCT116 cancer cell lines with CRISPR-Cas9”
From Frequent loss of heterozygosity in CRISPR-Cas9–edited early human embryos (2021) · cites this paper as [16]
In the paper’s results (page unavailable), 10 of 155 ZNF516-targeted HCT116 clones (6%) had loss of heterozygosity downstream of ZNF516, which the authors used to assess chromosome 18q arm truncation. Our copy does not state that the HCT116 cells were p53-competent, so that qualifier is unsupported. (LLM, conf=0.86)
Experimental — accuracy not yet measuredThe proximity of the CRISPR-Cas9-targeted locus to the telomere significantly increases the possibility of inadvertent chromosome arm truncation.
“For example, Przewrocka et al. demonstrate that the proximity of the CRISPR-Cas9-targeted locus to the telomere significantly increases the possibility of inadvertent chromosome arm truncation (16).”
From Frequent loss of heterozygosity in CRISPR-Cas9–edited early human embryos (2021) · cites this paper as [16]
The paper reports arm truncation after targeting the telomere-proximal ZNF516 gene, but not after targeting the more distant MEX3C gene. Its discussion says proximity to telomeres *could* elevate arm deletions; it does not establish that proximity significantly increases their likelihood. (LLM, conf=0.87)
Experimental — accuracy not yet measured
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Citations
Cites
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Research works printed but not linked: 6
- Jinek, 2013, RNA-programmed genome editing in human cells, eLife, 2013, 1
- Cong, 2013, Multiplex genome engineering using CRISPR/Cas systems, Science, 339, 819, 10.1126/science.1231143 · DOI · Paper page
- Mali, 2013, RNA-guided human genome engineering via Cas9, Science, 339, 823, 10.1126/science.1232033 · DOI · Paper page
- Kosicki, 2018, Repair of double-strand breaks induced by CRISPR-Cas9 leads to large deletions and complex rearrangements, Nat Biotechnol, 36, 765, 10.1038/nbt.4192 · DOI · Paper page
- Rayner, 2019, CRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methods, CRISPR J, 2, 406, 10.1089/crispr.2019.0006 · DOI
- Cullot, 2019, CRISPR-Cas9 genome editing induces megabase-scale chromosomal truncations, Nat Commun, 10, 1, 10.1038/s41467-019-09006-2 · DOI · Paper page
Cited by
Sorted by citations
Frequent loss of heterozygosity in CRISPR-Cas9–edited early human embryos
Alanis-Lobato, G., Zohren, J., McCarthy, A., Fogarty, N. M. E., Kubikova, N., Hardman, E., Greco, M., Wells, D., Turner, J. M. A., & Niakan, K. K. (2021) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2004832117 · Cited by 234
Metadata
- Authors
- Przewrocka, J., Rowan, A., Rosenthal, R., Kanu, N., & Swanton, C.
- Affiliations
- The Francis Crick Institute (GB)
- University College London (GB)
- Corresponding author
- Swanton, C.
- Venue
- Annals of Oncology
- Publisher
- Elsevier BV
- Published
- 2020-09
- Volume
- 31
- Issue
- 9
- Pages
- 1270-1273
- ISSN
- 0923-7534 (ISSN-L)
- PMC
- PMC7487774
- OpenAlex
- W3025068296
- Cited by
- 33
- References
- 0
- Citations by year
- 2025: 2 · 2024: 10 · 2023: 6 · 2022: 6 · 2021: 8 · 2020: 1
- FWCI
- 1.70
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Annals of Oncology (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- UCL Discovery (University College London) (submittedVersion)
- Figshare (submittedVersion, cc-by)
- Funders
- Wellcome Trust (FC001169)
- Francis Crick Institute (10169, 10002)
- Cancer Research UK (FC001169)
- 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
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chromatin Remodeling and Cancer
- Keywords
- CRISPR/Cas9, off-target effects, genome editing, clone screening, cancer therapy
- MeSH terms
- Humans, Gene Targeting, Chromosomal Instability, CRISPR-Cas Systems
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