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Low Incidence of Off-Target Mutations in Individual CRISPR-Cas9 and TALEN Targeted Human Stem Cell Clones Detected by Whole-Genome Sequencing
Open accessAbstract
Genome editing has attracted wide interest for the generation of cellular models of disease using human pluripotent stem cells and other cell types. CRISPR-Cas systems and TALENs can target desired genomic sites with high efficiency in human cells, but recent publications have led to concern about the extent to which these tools may cause off-target mutagenic effects that could potentially confound disease-modeling studies. Using CRISPR-Cas9 and TALEN targeted human pluripotent stem cell clones, we performed whole-genome sequencing at high coverage in order to assess the degree of mutagenesis across the entire genome. In both types of clones, we found that off-target mutations attributable to the nucleases were very rare. From this analysis, we suggest that, although some cell types may be at risk for off-target mutations, the incidence of such effects in human pluripotent stem cells may be sufficiently low and thus not a significant concern for disease modeling and other applications.
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“To further complicate this issue, reprogramming and genetic engineering itself have also been implicated in increasing genomic variation.169–171”
From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [170]
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Metadata
- Authors
- Veres, A., Gosis, B. S., Ding, Q., Collins, R., Ragavendran, A., Brand, H., Erdin, S., Cowan, C. A., Talkowski, M. E., & Musunuru, K.
- Affiliations
- Harvard University (US)
- Massachusetts General Hospital (US)
- Broad Institute (US)
- Corresponding author
- Musunuru, Kiran
- ORCID
- Venue
- Cell Stem Cell
- Publisher
- Elsevier BV
- Published
- 2014-07
- Volume
- 15
- Issue
- 1
- Pages
- 27-30
- ISSN
- 1875-9777 (ISSN-L)
- 1934-5909
- PMC
- PMC4082799
- OpenAlex
- W1974343902
- Cited by
- 474
- References
- 32
- Citations by year
- 2026: 6 · 2025: 8 · 2024: 10 · 2023: 24 · 2022: 24 · 2021: 32 · 2020: 33 · 2019: 43 · 2018: 46 · 2017: 61 · 2016: 72 · 2015: 86 · 2014: 29
- FWCI
- 33.92
- Type
- journal article
- Language
- en
- Update notices
- erratum (2014-08-07)
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell stem cell (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
- Funders
- National Heart, Lung, and Blood Institute (U01HL100408, R00HL098364, R01HL118744)
- National Institute of Mental Health (R00MH095867)
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK097768)
- 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 › Pluripotent Stem Cells Research
- Keywords
- off-target mutations, CRISPR/Cas9, TALEN, genome editing, human pluripotent stem cells, mutagenesis, disease modeling, whole genome sequencing
- MeSH terms
- Sortilin, Base Sequence, Clone Cells, Endonucleases, Genetic Engineering, Humans, Molecular Sequence Data, Mutation, Organ Specificity, Incidence, Genome, Sequence Analysis, DNA, Adaptor Proteins, Vesicular Transport, Pluripotent Stem Cells, CRISPR-Cas Systems
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