Whole-genome sequencing analysis reveals high specificity of CRISPR/Cas9 and TALEN-based genome editing in human iPSCs
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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 Smith et al., 2014
Incorporation of MP/bridged nucleic acids/locked nucleic acids increases on-target efficiency and decreases off-target effects.
“Incorporation of MP/bridged nucleic acids/locked nucleic acids | Increased on-target efficiency and decreased off-target effects”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as [78]
The paper’s main text reports efficient, highly specific CRISPR/Cas9 and TALEN editing in human iPSCs, but does not attribute those outcomes to incorporation of modified nucleic acids. Searches for MP, bridged or locked nucleic acids, and chemically modified guide RNA found no passage addressing their effects on on-target efficiency or off-target activity; page numbers were unavailable in our copy. (LLM, conf=0.76)
Experimental — accuracy not yet measuredChemical modification of gRNA by replacing RNA nucleotides with DNA nucleotides increases on-target efficiency and decreases off-target effects.
“| gRNA improvement | Chemical modifications [76–78] | Replacement of RNA nucleotides into DNA nucleotides | Increased on-target efficiency and decreased off-target effects |”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as [78]
The paper’s discussion of whole-genome sequencing in edited human iPSCs reports efficient, highly specific Cas9 and TALEN editing, but does not address chemically modified gRNAs. Searches for replacing RNA nucleotides with DNA nucleotides and for effects of gRNA modification on on-target efficiency or off-target activity found no relevant passage in our copy. (LLM, conf=0.76)
Experimental — accuracy not yet measured
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BackgroundOff‐target effects of engineered nucleases
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- Authors
- Smith, C., Gore, A., Yan, W., Abalde-Atristain, L., Li, Z., He, C., Wang, Y., Brodsky, R. A., Zhang, K., Cheng, L., & Ye, Z.
- Affiliations
- Johns Hopkins University (US)
- University of California San Diego (US)
- Corresponding author
- Cheng, Linzhao
- Ye, Zhaohui
- ORCID
- Venue
- Cell stem cell
- Publisher
- Elsevier BV
- Published
- 2014-07
- Volume
- 15
- Issue
- 1
- Pages
- 12-13
- ISSN
- 1875-9777 (ISSN-L)
- 1934-5909
- PMC
- PMC4338993
- OpenAlex
- W2028030005
- Cited by
- 342
- References
- 10
- Citations by year
- 2026: 9 · 2025: 4 · 2024: 8 · 2023: 14 · 2022: 16 · 2021: 25 · 2020: 17 · 2019: 29 · 2018: 28 · 2017: 40 · 2016: 56 · 2015: 71 · 2014: 25
- FWCI
- 26.26
- 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 Institutes of Health (5r01hl073781-10)
- National Heart, Lung, and Blood Institute (R01 HL-073781, U01HL107446, 2R01 HL073781, T32HL007525)
- National Cancer Institute (P30CA006973)
- National Institute of General Medical Sciences (T32GM007814)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- CRISPR/Cas9, TALEN, genome editing specificity, human induced pluripotent stem cells, zinc finger nucleases, genome editing efficiency, off-target effects
- MeSH terms
- Cell Line, DNA Repair, Endonucleases, Genetic Engineering, Humans, Genetic Therapy, Zinc Fingers, Genome, Sequence Analysis, DNA, Induced Pluripotent Stem Cells, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems
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