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

Abstract:
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.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1007/s00299-018-2355-9

Authors


More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Plant Sciences
Sub department:
Plant Sciences
Role:
Author
ORCID:
0000-0002-0215-3209
More by this author
Role:
Author
ORCID:
0000-0001-9386-1683


Publisher:
Springer Nature
Journal:
Plant Cell Reports More from this journal
Volume:
38
Issue:
4
Pages:
437-441
Place of publication:
Germany
Publication date:
2018-11-13
Acceptance date:
2018-11-07
DOI:
EISSN:
1432-203X
ISSN:
1432-203X
Pmid:
30426198


Language:
English
Keywords:
Pubs id:
1139953
Local pid:
pubs:1139953
Deposit date:
2020-10-26

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