GE-WRKY52 grape lines were generated by CRISPR-Cas editing of the transcription factor gene wrky52 in Vitis vinifera cultivar Thompson Seedless.
Transcription factors of the WRKY family have been shown to play roles in biotic stress responses.
Aim of the mutation of was the knockout of WRKY52 which enhances the resistance to Botrytis cinerea.
Four guide RNAs targeting the first exon region of the wrky52 gene were used for CRISPR-Cas editing. Transgenic plants were obtained via Agrobacterium-mediated transformation, using somatic embryos of Vitis vinifera.
Analysis of T0 transgenic plants verified 22 mutant plants; 15 lines carried biallelic mutations and seven were heterozygous. The majority of the mutations were smaller deletions but also large deletions were found in the mutant plants.
Sequencing of potential off-target sites for all four targets revealed no off-target events.
Knockout of WRKY52 in grape enhanced the resistance to Botrytis cinerea.
Source:
Wang et al. (2018), Plant Biotech J 16, 844–855; doi:10.1111/pbi.12832