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

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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, Lin, Ren-Jang, Yee, Jiing-Kuan·2015·Nature Biotechnology

DOI 10.1038/nbt.3127

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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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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
ORCID: Wang, Xiaoling
Wu, Xiwei
Wang, Jinhui
Wang, Yingjia
Qiu, Zhaojun
Huang, He
Lin, Ren-Jang
Venue: Nature Biotechnology
Publisher: Springer Science and Business Media LLC
Published: 2015-01-19
Volume: 33
Issue: 2
Pages: 175-178
ISSN: 0733-222X (ISSN-L)
1087-0156
1546-1696
DOI: 10.1038/nbt.3127
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
Licence: www.springer.com/tdm (tdm)
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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Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors

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