Cited by 323CRISPR and Genetic EngineeringRNA regulation and diseaseDNA and Nucleic Acid Chemistry

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Detection of on-target and off-target mutations generated by CRISPR/Cas9 and other sequence-specific nucleases

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Zischewski, Julia, Fischer, Rainer, Bortesi, Luisa·2017·Biotechnology Advances

DOI 10.1016/j.biotechadv.2016.12.003

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Abstract

The development of customizable sequence-specific nucleases such as TALENs, ZFNs and the powerful CRISPR/Cas9 system has revolutionized the field of genome editing. The CRISPR/Cas9 system is particularly versatile and has been applied in numerous species representing all branches of life. Regardless of the target organism, all researchers using sequence-specific nucleases face similar challenges: confirmation of the desired on-target mutation and the detection of off-target events. Here, we evaluate the most widely-used methods for the detection of on-target and off-target mutations in terms of workflow, sensitivity, strengths and weaknesses.

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Authors: Zischewski, J., Fischer, R., & Bortesi, L.
Affiliations: RWTH Aachen University (DE)
Fraunhofer Institute for Molecular Biology and Applied Ecology (DE)
Corresponding author: Bortesi, Luisa
ORCID: Zischewski, Julia
Fischer, Rainer
Bortesi, Luisa
Venue: Biotechnology Advances
Publisher: Elsevier BV
Published: 2017-01
Volume: 35
Issue: 1
Pages: 95-104
ISSN: 0734-9750 (ISSN-L)
DOI: 10.1016/j.biotechadv.2016.12.003
OpenAlex: W2565555627
Cited by: 323
References: 78
Citations by year: 2026: 14 · 2025: 29 · 2024: 32 · 2023: 31 · 2022: 35 · 2021: 41 · 2020: 40 · 2019: 46 · 2018: 37 · 2017: 18
FWCI: 3.04
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: cc-by-nc-nd
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
creativecommons.org/licenses/by-nc-nd/4.0/ (vor)
Locations: Biotechnology Advances (publishedVersion, cc-by-nc-nd)
PubMed (publishedVersion)
Research Publications (Maastricht University) (publishedVersion)
RWTH Publications (RWTH Aachen) (publishedVersion, other-oa)
Fraunhofer-Publica (Fraunhofer-Gesellschaft) (submittedVersion)
Funders: European Research Council (269110)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
Keywords: CRISPR/Cas9, off-target mutations, TALEN, zinc finger nucleases, sequence-specific nucleases, genome editing, sensitivity
MeSH terms: Gene Editing, Animals, Base Sequence, Cell Line, Deoxyribonucleases, Humans, Mutation, Mice, CRISPR-Cas Systems

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Detection of on-target and off-target mutations generated by CRISPR/Cas9 and other sequence-specific nucleases

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