CRISPR-Cas9-Mediated Targeted Mutagenesis in Seedless Grapes
October 18, 2017| |
The CRISPR-Cas9 system is a powerful tool for editing plant genomes. Genome editing of grape (Vitis vinifera) suspension cells using the CRISPR-Cas9 system has been demonstrated. However, it has not been established whether this system can be applied to get biallelic mutations in the first generation of grape.
Xianhang Wang of the Northwest A&F University in China designed four guide RNAs for the VvWRKY52 transcription factor gene using the CRISPR-Cas9 system on somatic embryos of the Thompson Seedless cultivar. Analysis of the first generation mutant plants verified 22 mutants, 15 of which carried biallelic mutations and 7 were heterozygous.
A range of editing events, including large deletions, was found in the mutant plants, while smaller deletions comprised the majority of the detected mutations. Analysis also revealed no off-target mutation occurred. Knock out of VvWRKY52 gene in grape increased the plant's resistance to Botrytis cinerea.
The CRISPR/Cas9 system allows precise genome editing in the first generation of grape and represents a useful tool for gene functional analysis and grape molecular breeding.
For more on this study, read the article in Plant Biotechnology Journal.
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