Cited by 1,137CRISPR and Genetic EngineeringDNA Repair MechanismsRNA regulation and disease

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Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining

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Maruyama, Takeshi, Dougan, Stephanie K, Truttmann, Matthias C, Bilate, Angelina M, Ingram, Jessica R, Ploegh, Hidde L·2015·Nature Biotechnology

DOI 10.1038/nbt.3190PMC PMC4618510

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Abstract

Methods to introduce targeted double-strand breaks (DSBs) into DNA enable precise genome editing by increasing the rate at which externally supplied DNA fragments are incorporated into the genome through homologous recombination. The efficiency of these methods is limited by nonhomologous end joining (NHEJ), an alternative DNA repair pathway that competes with homology-directed repair (HDR). To promote HDR at the expense of NHEJ, we targeted DNA ligase IV, a key enzyme in the NHEJ pathway, using the inhibitor Scr7. Scr7 treatment increased the efficiency of HDR-mediated genome editing, using Cas9 in mammalian cell lines and in mice for all four genes examined, up to 19-fold. This approach should be applicable to other customizable endonucleases, such as zinc finger nucleases and transcription activator-like effector nucleases, and to nonmammalian cells with sufficiently conserved mechanisms of NHEJ and HDR.

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Authors: Maruyama, T., Dougan, S. K., Truttmann, M. C., Bilate, A. M., Ingram, J. R., & Ploegh, H. L.
Affiliations: Whitehead Institute for Biomedical Research (US)
Corresponding author: Ploegh, Hidde L
ORCID: Maruyama, Takeshi
Dougan, Stephanie K
Truttmann, Matthias C
Bilate, Angelina M
Ingram, Jessica R
Ploegh, Hidde L
Venue: Nature Biotechnology
Publisher: Springer Science and Business Media LLC
Published: 2015-03-23
Volume: 33
Issue: 5
Pages: 538-542
ISSN: 0733-222X (ISSN-L)
1087-0156
1546-1696
DOI: 10.1038/nbt.3190
PMC: PMC4618510
OpenAlex: W2008147758
Cited by: 1,137
References: 32
Citations by year: 2026: 39 · 2025: 58 · 2024: 62 · 2023: 67 · 2022: 108 · 2021: 101 · 2020: 117 · 2019: 122 · 2018: 125 · 2017: 139 · 2016: 143 · 2015: 55
FWCI: 66.47
Type: journal article
Language: en
Update notices: erratum (2016-02-05)
Open access: green
www.ncbi.nlm.nih.gov
Licence: www.springer.com/tdm (tdm)
www.springer.com/tdm (vor)
Locations: Nature Biotechnology (publishedVersion)
PubMed (publishedVersion)
PubMed Central (acceptedVersion)
DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc)
DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc-sa)
PMC
Funders: National Institute of Allergy and Infectious Diseases (R01AI087879)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
Keywords: CRISPR/Cas9, genome editing efficiency, non-homologous end joining, homology-directed repair, DNA ligase IV, DNA double-strand break repair, zinc finger nucleases, TALEN
MeSH terms: DNA Ligase ATP, Animals, Cell Line, DNA Repair, Genetic Engineering, DNA Ligases, Pyrimidines, Schiff Bases, Zinc Fingers, Genome, Mice, DNA Breaks, Double-Stranded, Homologous Recombination, DNA End-Joining Repair, CRISPR-Cas Systems

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Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining

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