Cited by 442CRISPR and Genetic EngineeringVirus-based gene therapy researchCAR-T cell therapy research

Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors

Wang, Xiaoling, Wang, Yebo, Wu, Xiwei, Wang, Jinhui, Wang, Yingjia, Qiu, Zhaojun, Chang, Tammy, Huang, He +2 more·2015·Nature Biotechnology

Abstract

The utility of CRISPR-Cas9 and TALENs for genome editing may be compromised by their off-target activity. We show that integrase-defective lentiviral vectors (IDLVs) can detect such off-target cleavage with a frequency as low as 1%. In the case of Cas9, we find frequent off-target sites with a one-base bulge or up to 13 mismatches between the single guide RNA (sgRNA) and its genomic target, which refines sgRNA design.

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Crossref · 10.1038/nbt.3127

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20 of 20 references verified0 citation errors · 0 not checkable

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  • IDLV displays a propensity to integrate into the vicinity of DSBs.

    “One such labeling tool is called the integrase-defective lentiviral vectors (IDLV), which displays the propensity to integrate into the vicinity of DSBs (Wang et al., 2015; Kim et al., 2019).”

    From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as Wang et al., 2015

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Authors
Wang, X., Wang, Y., Wu, X., Wang, J., Wang, Y., Qiu, Z., Chang, T., Huang, H., Lin, R.-J., & Yee, J.-K.
Affiliations
  • City Of Hope National Medical Center (US)
  • First Affiliated Hospital Zhejiang University (CN)
Corresponding author
Yee, Jiing-Kuan
Venue
Nature Biotechnology
Published
2015-01-19
Volume
33
Issue
2
Pages
175-178
ISSN
  • 0733-222X (ISSN-L)
  • 1087-0156
  • 1546-1696
OpenAlex
W2051255304
Cited by
442
References
20
Citations by year
2026: 5 · 2025: 17 · 2024: 18 · 2023: 25 · 2022: 31 · 2021: 38 · 2020: 47 · 2019: 36 · 2018: 46 · 2017: 57 · 2016: 70 · 2015: 52
FWCI
32.55
Type
journal article
Language
en
Open access
closed
Locations
  • Nature Biotechnology (publishedVersion)
  • PubMed (publishedVersion)
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
  • Health Sciences › Medicine › Oncology › CAR-T cell therapy research
Keywords
off-target cleavage, CRISPR/Cas9, TALEN, integrase-defective lentiviral vectors, genome editing, single guide RNA design, mismatch tolerance
MeSH terms
Genetic Vectors, Humans, Genome, Human, Lentivirus, RNA Editing, Integrases, HEK293 Cells, High-Throughput Nucleotide Sequencing, CRISPR-Cas Systems

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