2018
DOI: 10.1007/s00299-018-2355-9
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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 s… Show more

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Cited by 111 publications
(79 citation statements)
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References 38 publications
(46 reference statements)
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“…The HRFA of the selected lines indicated no insertions or deletions in StPPO1 and StPPO4 genes. The presence of multiple mismatches into the seed region (defined as the 8-12 nt proximal to the PAM) between the selected sgRNAs and the rest of StPPO genes could explain their specificity for StPPO2 (Hahn and Nekrasov, 2019). On the other hand, the use of RNPs as delivery method for the CRISPR/Cas9 system has been proposed to reduce the incidence of off targets effects, due to the rapid degradation of the Cas9 nuclease and the sgRNAs in the cell (Nadakuduti et al, 2018;Zhang et al, 2018a;Hahn and Nekrasov, 2019).…”
Section: Discussionmentioning
confidence: 99%
See 1 more Smart Citation
“…The HRFA of the selected lines indicated no insertions or deletions in StPPO1 and StPPO4 genes. The presence of multiple mismatches into the seed region (defined as the 8-12 nt proximal to the PAM) between the selected sgRNAs and the rest of StPPO genes could explain their specificity for StPPO2 (Hahn and Nekrasov, 2019). On the other hand, the use of RNPs as delivery method for the CRISPR/Cas9 system has been proposed to reduce the incidence of off targets effects, due to the rapid degradation of the Cas9 nuclease and the sgRNAs in the cell (Nadakuduti et al, 2018;Zhang et al, 2018a;Hahn and Nekrasov, 2019).…”
Section: Discussionmentioning
confidence: 99%
“…In order to avoid inducing mutations in other StPPO genes, the two selected sgRNAs were analyzed for possible off target activity. Considering up to four mismatches (Hahn and Nekrasov, 2019) StPPO1 and StPPO4 genes were identified as possible off targets of sgRNA564 ( Figure 1B and Supplementary Figures S2 and S3). Four mismatches at positions +1, +2, +8, and +13 from the Protospacer Adjacent Motif (PAM) were identified in the potential off target site on StPPO1 and four mismatches at positions +1, +8, +13, and +20, in the potential off target site on StPPO4 ( Figure 1B).…”
Section: Sgrna Design On Stppo2 Gene and Off Target Predictionmentioning
confidence: 99%
“…Transgene-free genome editing is the preferred choice for the application of cis-engineering for crop improvement and commercialization of genome-edited crops. Genome editing with stable expression of CRISPR/Cas DNA involves the integration of the construct into the host genome, raising concerns about undesirable offtarget changes and biosecurity 60,111 . Genetic segregation by selfing or crossing can be used to obtain transgene-free edited plants.…”
Section: Transgene-free Genome Editingmentioning
confidence: 99%
“…Transformation and tissue culture are crucial for CRISPR-mediated genome editing, while their efficiencies are challenged for most of crop plants. Last but not the least, off-targeting is still an intensive concern for plant scientists, though numerous studies have evidenced the precision of CRISPR-mediated genome editing in plants [170]. All of these challenges should be addressed in the future studies in order to promote the applications of CRISPR systems in crop improvement.…”
Section: Perspectivesmentioning
confidence: 99%