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• W2895564510 abstract "The CRISPR/Cas9 system has been extensively applied for crop improvement. However, our understanding of Cas9 specificity is very limited in Cas9-edited plants. To identify on- and off-target mutation in an edited crop, we described whole genome sequencing (WGS) of 14 Cas9-edited cotton plants targeted to three genes, and three negative (Ne) control and three wild-type (WT) plants. In total, 4188–6404 unique single-nucleotide polymorphisms (SNPs) and 312–745 insertions/deletions (indels) were detected in 14 Cas9-edited plants compared to WT, negative and cotton reference genome sequences. Since the majority of these variations lack a protospacer-adjacent motif (PAM), we demonstrated that the most variations following Cas9-edited are due either to somaclonal variation or/and pre-existing/inherent variation from maternal plants, but not off-target effects. Of a total of 4413 potential off-target sites (allowing ≤5 mismatches within the 20-bp sgRNA and 3-bp PAM sequences), the WGS data revealed that only four are bona fide off-target indel mutations, validated by Sanger sequencing. Moreover, inherent genetic variation of WT can generate novel off-target sites and destroy PAMs, which suggested great care should be taken to design sgRNA for the minimizing of off-target effect. These findings suggested that CRISPR/Cas9 system is highly specific for cotton plants." @default.
• W2895564510 created "2018-10-12" @default.
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• W2895564510 date "2018-10-30" @default.
• W2895564510 modified "2023-10-16" @default.
• W2895564510 title "Whole genome sequencing reveals rare off‐target mutations and considerable inherent genetic or/and somaclonal variations in <scp>CRISPR</scp> /Cas9‐edited cotton plants" @default.
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• W2895564510 doi "https://doi.org/10.1111/pbi.13020" @default.
• W2895564510 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6587709" @default.
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• W2895564510 hasPublicationYear "2018" @default.
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