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

: Zischewski, J.; Fischer, R.; Bortesi, L.

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Biotechnology Advances 35 (2017), No.1, pp.95-104
ISSN: 0734-9750
ISSN: 1873-1899
Journal Article, Electronic Publication
Fraunhofer IME ()

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.