Large deletions induced by Cas9 cleavage
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Adikusama et al. detected large deletions between 100bp and 2.3kb in 45% of 127 early mouse embryos using long-range PCR.
“Adikusama et al. targeted six genes in a total of 127 early mouse embryos and detected large deletions (between 100bp and 2.3kb) in 45% of their samples using long-range PCR (10).”
From Frequent loss of heterozygosity in CRISPR-Cas9–edited early human embryos (2021) · cites this paper as [10]
The 'Large deletions induced by Cas9 cleavage' section reports deletions over 100 bp in 57 of 127 samples (45%) using larger PCR products, with a maximum deletion of 2.3 kb. The 'Sample size' section identifies six targeted loci but calls the 127 subjects 'founder embryos/mice,' not exclusively early mouse embryos. (LLM, conf=0.88)
Experimental — accuracy not yet measuredCRISPR-Cas9-induced deletions larger than the PCR fragment can eliminate one or both primer annealing sites, leading to amplification of only one allele and a false impression of unsuccessful targeting or a single homozygous event.
“CRISPR-Cas9-induced deletions larger than these fragments in either direction would eliminate one or both PCR primer annealing sites.This in turn can lead to amplification of only one allele, giving the false impression that targeting was unsuccessful or that there is a single homozygous event at the on-target site (9,10,15).”
From Frequent loss of heterozygosity in CRISPR-Cas9–edited early human embryos (2021) · cites this paper as [10]
Large deletions were generally more prevalent when targeting intronic regions (>70%) than exons (20%).
“Of note, large deletions were generally more prevalent when they targeted intronic regions (>70%) than when they targeted exons (20%).”
From Frequent loss of heterozygosity in CRISPR-Cas9–edited early human embryos (2021) · cites this paper as [10]
The “Large deletions induced by Cas9 cleavage” section reports large-deletion frequencies, but does not compare intronic and exonic targets. Searches for intronic and exonic targeting and the stated percentages found no passage supporting the >70% versus 20% comparison; the copy’s mention of approximately 20% concerns edited mouse embryonic stem cells, not exons. (LLM, conf=0.72)
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- Authors
- Adikusuma, F., Piltz, S., Corbett, M. A., Turvey, M., McColl, S. R., Helbig, K. J., Beard, M. R., Hughes, J., Pomerantz, R. T., & Thomas, P. Q.
- Affiliations
- Diponegoro University (ID)
- South Australian Health and Medical Research Institute (AU)
- The University of Adelaide (AU)
- Singapore-MIT Alliance for Research and Technology (SG)
- La Trobe University (AU)
- Temple University (US)
- Corresponding author
- Thomas, Paul Q.
- ORCID
- Venue
- Nature
- Published
- 2018-08
- Volume
- 560
- Issue
- 7717
- Pages
- E8-E9
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- OpenAlex
- W2885661865
- Cited by
- 371
- References
- 8
- Citations by year
- 2026: 19 · 2025: 39 · 2024: 63 · 2023: 60 · 2022: 69 · 2021: 44 · 2020: 41 · 2019: 26 · 2018: 10
- FWCI
- 10.76
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Funders
- Oregon Health and Science University
- Leona M. and Harry B. Helmsley Charitable Trust
- South Australian Health and Medical Research Institute
- Temple University
- Consorzio Interuniversitario Biotecnologie
- Moxie Foundation
- La Trobe University
- University of Adelaide
- Government of Jiangxi Province
- Universitas Diponegoro
- Institute for Basic Science (IBSR021-D1, R021-D1)
- Directorate for Biological Sciences
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Keywords
- CRISPR/Cas9 genome editing, MYBPC3 mutation, large deletions, DNA double-strand break repair, mouse embryo model
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