Occurrence and Nature of Off-Target Modifications by CRISPR-Cas Genome Editing in Plants

Mark H.J. Sturme*, Jan P. van der Berg, Lianne M.S. Bouwman, Adinda De Schrijver, Ruud A. de Maagd, Gijs A. Kleter, Evy Battaglia-De Wilde

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

21 Citations (Scopus)

Abstract

CRISPR-Cas-based genome editing allows for precise and targeted genetic modification of plants. Nevertheless, unintended off-target edits can arise that might confer risks when present in gene-edited food crops. Through an extensive literature review we gathered information on CRISPR-Cas off-target edits in plants. Most observed off-target changes were small insertions or deletions (1-22 bp) or nucleotide substitutions, and large deletions (>100 bp) were rare. One study detected the insertion of vector-derived DNA sequences, which is important considering the risk assessment of gene-edited plants. Off-target sites had few mismatches (1-3 nt) with the target sequence and were mainly located in protein-coding regions, often in target gene homologues. Off-targets edits were predominantly detected via biased analysis of predicted off-target sites instead of unbiased genome-wide analysis. CRISPR-Cas-edited plants showed lower off-target mutation frequencies than conventionally bred plants. This Review can aid discussions on the relevance of evaluating off-target modifications for risk assessment of CRISPR-Cas-edited plants.

Original languageEnglish
Pages (from-to)192-201
JournalACS Agricultural Science and Technology
Volume2
Issue number2
DOIs
Publication statusPublished - 18 Apr 2022

Keywords

  • CRISPR-Cas
  • genome editing
  • off-target modifications
  • plants

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