Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors - PubMed (original) (raw)
doi: 10.1038/nbt.3127. Epub 2015 Jan 19.
Affiliations
- PMID: 25599175
- DOI: 10.1038/nbt.3127
Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors
Xiaoling Wang et al. Nat Biotechnol. 2015 Feb.
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.
Comment in
- Mapping the precision of genome editing.
Gabriel R, von Kalle C, Schmidt M. Gabriel R, et al. Nat Biotechnol. 2015 Feb;33(2):150-2. doi: 10.1038/nbt.3142. Nat Biotechnol. 2015. PMID: 25658281 No abstract available.
Similar articles
- Detection of CRISPR/Cas9-Generated Off-Target Effect by Integration-Defective Lentiviral Vector.
Wang X, Wu Y, Yee JK. Wang X, et al. Methods Mol Biol. 2021;2162:243-260. doi: 10.1007/978-1-0716-0687-2_14. Methods Mol Biol. 2021. PMID: 32926387 - A Protocol for the Production of Integrase-deficient Lentiviral Vectors for CRISPR/Cas9-mediated Gene Knockout in Dividing Cells.
Vijayraghavan S, Kantor B. Vijayraghavan S, et al. J Vis Exp. 2017 Dec 12;(130):56915. doi: 10.3791/56915. J Vis Exp. 2017. PMID: 29286484 Free PMC article. - Systematic analysis of CRISPR-Cas9 mismatch tolerance reveals low levels of off-target activity.
Anderson EM, Haupt A, Schiel JA, Chou E, Machado HB, Strezoska Ž, Lenger S, McClelland S, Birmingham A, Vermeulen A, Smith Av. Anderson EM, et al. J Biotechnol. 2015 Oct 10;211:56-65. doi: 10.1016/j.jbiotec.2015.06.427. Epub 2015 Jul 17. J Biotechnol. 2015. PMID: 26189696 - Lentiviral Vectors for Delivery of Gene-Editing Systems Based on CRISPR/Cas: Current State and Perspectives.
Dong W, Kantor B. Dong W, et al. Viruses. 2021 Jul 1;13(7):1288. doi: 10.3390/v13071288. Viruses. 2021. PMID: 34372494 Free PMC article. Review. - 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. Martin F, et al. Int J Mol Sci. 2016 Sep 8;17(9):1507. doi: 10.3390/ijms17091507. Int J Mol Sci. 2016. PMID: 27618019 Free PMC article. Review.
Cited by
- The Evolution of Nucleic Acid-Based Diagnosis Methods from the (pre-)CRISPR to CRISPR era and the Associated Machine/Deep Learning Approaches in Relevant RNA Design.
Chakraborty SS, Ray Dutta J, Ganesan R, Minary P. Chakraborty SS, et al. Methods Mol Biol. 2025;2847:241-300. doi: 10.1007/978-1-0716-4079-1_17. Methods Mol Biol. 2025. PMID: 39312149 - Cell-Penetrating Peptides and CRISPR-Cas9: A Combined Strategy for Human Genetic Disease Therapy.
de Morais CCPL, Correia EM, Bonamino MH, Vasconcelos ZFM. de Morais CCPL, et al. Hum Gene Ther. 2024 Oct;35(19-20):781-797. doi: 10.1089/hum.2024.020. Hum Gene Ther. 2024. PMID: 39276086 Review. - Precision in Action: The Role of Clustered Regularly Interspaced Short Palindromic Repeats/Cas in Gene Therapies.
Banda A, Impomeni O, Singh A, Baloch AR, Hu W, Jaijyan DK. Banda A, et al. Vaccines (Basel). 2024 Jun 7;12(6):636. doi: 10.3390/vaccines12060636. Vaccines (Basel). 2024. PMID: 38932365 Free PMC article. Review. - CRISPR-Cas and CRISPR-based screening system for precise gene editing and targeted cancer therapy.
Qin M, Deng C, Wen L, Luo G, Meng Y. Qin M, et al. J Transl Med. 2024 May 30;22(1):516. doi: 10.1186/s12967-024-05235-2. J Transl Med. 2024. PMID: 38816739 Free PMC article. Review. - Beyond the promise: evaluating and mitigating off-target effects in CRISPR gene editing for safer therapeutics.
Lopes R, Prasad MK. Lopes R, et al. Front Bioeng Biotechnol. 2024 Jan 18;11:1339189. doi: 10.3389/fbioe.2023.1339189. eCollection 2023. Front Bioeng Biotechnol. 2024. PMID: 38390600 Free PMC article. Review.
References
- Nat Methods. 2013 Oct;10(10):973-6 - PubMed
- PLoS One. 2014 Apr 01;9(4):e93575 - PubMed
- Cell. 2013 Sep 12;154(6):1380-9 - PubMed
- Mol Ther. 2013 Jun;21(6):1151-9 - PubMed
- Nat Biotechnol. 2013 Sep;31(9):833-8 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources