Off-target mutations are rare in Cas9-modified mice
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WGS can directly uncover desired and unwanted editing events by comparing genome sequences before and after CRISPR/Cas9 editing.
“By comparing the genome sequences before and after CRISPR/Cas9 editing, WGS can directly uncover desired and unwanted editing events.”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as Iyer et al., 2015
Cites
Wang, H., Yang, H., Shivalila, C. S., Dawlaty, M. M., Cheng, A. W., Zhang, F., & Jaenisch, R. (2013) · Cell · doi:10.1016/j.cell.2013.04.025 · Cited by 3,495 · see citing works
Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity
Ran, F. A., Hsu, P. D., Lin, C.-Y., Gootenberg, J. S., Konermann, S., Trevino, A. E., Scott, D. A., Inoue, A., Matoba, S., Zhang, Y., et al. (2013) · Cell · doi:10.1016/j.cell.2013.08.021 · Cited by 3,299 · see citing works
Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects
Shen, B., Zhang, W., Zhang, J., Zhou, J., Wang, J.-Y., Chen, L., Wang, L., Hodgkins, A., Iyer, V., Huang, X., et al. (2014) · Nature Methods · doi:10.1038/nmeth.2857 · Cited by 827 · see citing works
Self-citationVeres, A., Gosis, B. S., Ding, Q., Collins, R., Ragavendran, A., Brand, H., Erdin, S., Cowan, C. A., Talkowski, M. E., & Musunuru, K. (2014) · Cell Stem Cell · doi:10.1016/j.stem.2014.04.020 · Cited by 474 · see citing works
Smith, C., Gore, A., Yan, W., Abalde-Atristain, L., Li, Z., He, C., Wang, Y., Brodsky, R. A., Zhang, K., Cheng, L., et al. (2014) · Cell stem cell · doi:10.1016/j.stem.2014.06.011 · Cited by 342 · see citing works
Suzuki, K., Yu, C., Qu, J., Li, M., Yao, X., Yuan, T., Goebl, A., Tang, S., Ren, R., Aizawa, E., et al. (2014) · Cell Stem Cell · doi:10.1016/j.stem.2014.06.016 · Cited by 169 · see citing works
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Haeussler, M., Schönig, K., Eckert, H., Eschstruth, A., Mianné, J., Renaud, J.-B., Schneider-Maunoury, S., Shkumatava, A., Teboul, L., Kent, J., et al. (2016) · Genome biology · doi:10.1186/s13059-016-1012-2 · Cited by 1,756 · see citing works
Off-target Effects in CRISPR/Cas9-mediated Genome Engineering
Zhang, X.-H., Tee, L. Y., Wang, X.-G., Huang, Q.-S., & Yang, S.-H. (2015) · Molecular Therapy - Nucleic Acids · doi:10.1038/mtna.2015.37 · Cited by 1,273 · see citing works
Off-target effects in CRISPR/Cas9 gene editing
Guo, C., Ma, X., Gao, F., & Guo, Y. (2023) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2023.1143157 · Cited by 625 · see citing works
“WGS analysis of off-target effects has been well documented in cell culture studies (Smith et al., 2014; Veres et al., 2014; Iyer et al., 2015).”
BackgroundZischewski, J., Fischer, R., & Bortesi, L. (2017) · Biotechnology Advances · doi:10.1016/j.biotechadv.2016.12.003 · Cited by 323 · see citing works
The Trem2 R47H variant confers loss-of-function-like phenotypes in Alzheimer's disease
Cheng-Hathaway, P. J., Reed-Geaghan, E. G., Jay, T. R., Casali, B. T., Bemiller, S. M., Puntambekar, S. S., von Saucken, V. E., Williams, R. Y., Karlo, J. C., Moutinho, M., et al. (2018) · Molecular Neurodegeneration · doi:10.1186/s13024-018-0262-8 · Cited by 252 · see citing works
Evaluating and enhancing target specificity of gene-editing nucleases and deaminases
Kim, D., Luk, K., Wolfe, S. A., & Kim, J.-S. (2019) · Annual Review of Biochemistry · doi:10.1146/annurev-biochem-013118-111730 · Cited by 171 · see citing works
Biased and unbiased methods for the detection of off-target cleavage by CRISPR/Cas9: An overview
Martin, F., Sánchez-Hernández, S., Gutiérrez-Guerrero, A., Pinedo-Gomez, J., & Benabdellah, K. (2016) · International Journal of Molecular Sciences · doi:10.3390/ijms17091507 · Cited by 92 · see citing works
“In theory, as the entire genome of a cell can now be sequenced, the WGS of edited cells could be used to study CRISPR/Cas9 specificity [36,37].”
Background
- Authors
- Iyer, V., Shen, B., Zhang, W., Hodgkins, A., Keane, T., Huang, X., & Skarnes, W. C.
- Affiliations
- Wellcome Sanger Institute (GB)
- Model Animal Research Center (CN)
- Corresponding author
- Huang, Xingxu
- Skarnes, William C
- Venue
- Nature Methods
- Published
- 2015-05-28
- Volume
- 12
- Issue
- 6
- Pages
- 479-479
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- OpenAlex
- W1555684022
- Cited by
- 185
- References
- 6
- Citations by year
- 2026: 4 · 2025: 4 · 2024: 15 · 2023: 7 · 2022: 10 · 2021: 11 · 2020: 12 · 2019: 20 · 2018: 36 · 2017: 27 · 2016: 28 · 2015: 11
- FWCI
- 13.41
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Wellcome Trust
- Medical Research Council (MR/L007428/1)
- Biotechnology and Biological Sciences Research Council (BB/M000281/1)
- National Key Research and Development Program of China (2011CB944301)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- off-target mutations, CRISPR/Cas9, genome editing, mouse model, genome editing specificity, off-target effects
- MeSH terms
- Animals, Breeding, Deoxyribonuclease I, Female, Male, Mice, Inbred C57BL, Mutation, RNA, Messenger, Gene Expression Regulation, Enzymologic, Sequence Alignment, Genome, Mice, Knockout, Mice, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Associated Proteins
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