Journal Article

Genome-wide specificity of plant genome editing by both CRISPR–Cas9 and TALEN

Mara, Kostlend, ANR-11-BTBR-0001,GENIUS,Ingénierie cellulaire : amélioration et innovation technologiques pour les plantes d'une agriculture, Collonnier, Cécile, Guyon-Debast, Anouchka, Charif, Delphine, Institut Jean-Pierre Bourgin - Sciences du végétal (IJPB) ; AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), ANR-17-EURE-0007,SPS-GSR,Ecole Universitaire de Recherche de Sciences des Plantes de Paris-Saclay, Charlot, Florence, Tepfer, Mark, Perroud, Pierre-François, Bessoltane, Nadia, Nogué, Fabien, CSIRO Health and Biosecurity [Australia] ; Commonwealth Scientific and Industrial Research Organisation [Australia] (CSIRO)

2022

Overview

Abstract CRISPR and TALENs are efficient systems for gene editing in many organisms including plants. In many cases the CRISPR–Cas or TALEN modules are expressed in the plant cell only transiently. Theoretically, transient expression of the editing modules should limit unexpected effects compared to stable transformation. However, very few studies have measured the off-target and unpredicted effects of editing strategies on the plant genome, and none of them have compared these two major editing systems. We conducted, in Physcomitrium patens , a comprehensive genome-wide investigation of off-target mutations using either a CRISPR–Cas9 or a TALEN strategy. We observed a similar number of differences for the two editing strategies compared to control non-transfected plants, with an average of 8.25 SNVs and 19.5 InDels for the CRISPR-edited plants, and an average of 17.5 SNVs and 32 InDels for the TALEN-edited plants. Interestingly, a comparable number of SNVs and InDels could be detected in the PEG-treated control plants. This shows that except for the on-target modifications, the gene editing tools used in this study did not show a significant off-target activity nor unpredicted effects on the genome, and did not lead to transgene integration. The PEG treatment, a well-established biotechnological method, in itself, was the main source of mutations found in the edited plants.

• Publisher: Nature Publishing Group,CCSD,Nature Publishing Group UK,Nature Portfolio
• Subject: 631/449 / 631/449/447 / 631/61/447/2311 / Biotechnology / CRISPR / Genetic transformation / Genome editing / Genomes / Humanities and Social Sciences / Life Sciences / multidisciplinary / Mutation / Science / Science (multidisciplinary) / Transcription activator-like effector nucleases
• ISBN: 0008066608000