CRISPR/Cas precision: do we need to worry about off-targeting in plants?

A - Papers appearing in refereed journals

Hahn, F. and Nekrasov, V. 2018. CRISPR/Cas precision: do we need to worry about off-targeting in plants? Plant Cell Reports.

AuthorsHahn, F. and Nekrasov, V.

The CRISPR/Cas technology has recently become the tool of choice for targeted genome modification in plants and beyond. Although CRSIPR/Cas offers a rapid and facile way of introducing changes at genomic loci of interest, its application is associated with off-targeting, i.e. introduction of unintended mutations at off-target sites within the genome, which has been reported frequently in the mammalian field. Here we summarise the current knowledge on the precision of CRISPR/Cas in plant systems and provide a summary of state-of the-art strategies for avoiding off-target mutations, as well as unintended on-target changes, in plants. These include using natural (e.g. Cas12a) or engineered (e.g. SpCas9-HF) CRISPR/Cas nucleases characterised by higher precision, as compared to the commonly used wild type SpCas9. In addition, we discuss the usage of CRISPR/Cas nucleases in the form of ribonucleoproteins (RNPs) as an option for reducing off-targeting in plants. Finally, we conclude that the most important factor for reducing CRISPR/Cas off-targeting remains careful selection of target sequences, for which we provide an overview of available online software tools and experimental guidance.

Year of Publication2018
JournalPlant Cell Reports
Digital Object Identifier (DOI)
PubMed ID30426198
Open accessPublished as ‘gold’ (paid) open access
FunderBBSRC Industrial Strategy Challenge
Funder project or codeDesigning Future Wheat (DFW) [ISPG]
Publisher's version
Output statusE-publication ahead of print
Publication dates
Online13 Nov 2018
Publication process dates
Accepted07 Nov 2018
Copyright licenseCC BY

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