Targeted Gene Correction Minimally Impacts Whole-Genome Mutational Load in Human-Disease-Specific Induced Pluripotent Stem Cell Clones
Open accessAbstract
The utility of genome editing technologies for disease modeling and developing cellular therapies has been extensively documented, but the impact of these technologies on mutational load at the whole-genome level remains unclear. We performed whole-genome sequencing to evaluate the mutational load at single-base resolution in individual gene-corrected human induced pluripotent stem cell (hiPSC) clones in three different disease models. In single-cell clones, gene correction by helper-dependent adenoviral vector (HDAdV) or Transcription Activator-Like Effector Nuclease (TALEN) exhibited few off-target effects and a low level of sequence variation, comparable to that accumulated in routine hiPSC culture. The sequence variants were randomly distributed and unique to individual clones. We also combined both technologies and developed a TALEN-HDAdV hybrid vector, which significantly increased gene-correction efficiency in hiPSCs. Therefore, with careful monitoring via whole-genome sequencing it is possible to apply genome editing to human pluripotent cells with minimal impact on genomic mutational load.
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- Authors
- Suzuki, K., Yu, C., Qu, J., Li, M., Yao, X., Yuan, T., Goebl, A., Tang, S., Ren, R., Aizawa, E., Zhang, F., Xu, X., Soligalla, R. D., Chen, F., Kim, J., Kim, N. Y., Liao, H.-K., Benner, C., Esteban, C. R., Jin, Y., Liu, G.-H., Li, Y., & Izpisua Belmonte, J. C.
- Affiliations
- Salk Institute for Biological Studies (US)
- BGI Group (China) (CN)
- Chinese Academy of Sciences (CN)
- Corresponding author
- Liu, Guang-Hui
- Li, Yingrui
- ORCID
- Venue
- Cell Stem Cell
- Publisher
- Elsevier BV
- Published
- 2014-07
- Volume
- 15
- Issue
- 1
- Pages
- 31-36
- ISSN
- 1875-9777 (ISSN-L)
- 1934-5909
- PMC
- PMC4144407
- OpenAlex
- W1974421923
- Cited by
- 169
- References
- 35
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 6 · 2023: 7 · 2022: 6 · 2021: 8 · 2020: 6 · 2019: 15 · 2018: 6 · 2017: 20 · 2016: 33 · 2015: 41 · 2014: 18
- FWCI
- 15.30
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell stem cell (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Heart, Lung, and Blood Institute (U01HL107442)
- National Cancer Institute (P30CA014195)
- 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 › Pluripotent Stem Cells Research
- Keywords
- induced pluripotent stem cells, gene correction, mutational load, off-target effects, helper-dependent adenoviral vectors, TALEN, single-cell clones, disease modeling, genome editing
- MeSH terms
- Adenoviridae, Clone Cells, DNA Repair, Endonucleases, Genetic Vectors, Humans, Mutation, Genetic Therapy, Genome, Sequence Analysis, DNA, Regenerative Medicine, Induced Pluripotent Stem Cells, HEK293 Cells, CRISPR-Cas Systems
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