title: CRISPR/Cas9-mediated efficient targeted mutagenesis in Chardonnay (Vitis vinifera L.).
abstract: The type II clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 system (CRISPR/Cas9) has been successfully applied to edit target genes in multiple plant species. However, it remains unknown whether this system can be used for genome editing in grape. In this study, we described genome editing and targeted gene mutation in 'Chardonnay' suspension cells and plants via the CRISPR/Cas9 system. Two single guide RNAs (sgRNAs) were designed to target distinct sites of the L-idonate dehydrogenase gene (IdnDH). CEL I endonuclease assay and sequencing results revealed the expected indel mutations at the target site, and a mutation frequency of 100% was observed in the transgenic cell mass (CM) as well as corresponding regenerated plants with expression of sgRNA1/Cas9. The majority of the detected mutations in transgenic CM were 1-bp insertions, followed by 1- to 3-nucleotide deletions. Off-target activities were also evaluated by sequencing the potential off-target sites, and no obvious off-target events were detected. Our results demonstrated that the CRISPR/Cas9 system is an efficient and specific tool for precise genome editing in grape.
authors: Beijing Key Laboratory of Grape Science and Enology and Key Laboratory of Plant Resource, Institute of Botany, the Chinese Academy of Sciences, Beijing 100093, PR China.|University of Chinese Academy of Sciences, Beijing 100049, PR China.
authors: Chong
authors: C
authors: Ren
authors: Beijing Key Laboratory of Grape Science and Enology and Key Laboratory of Plant Resource, Institute of Botany, the Chinese Academy of Sciences, Beijing 100093, PR China.|University of Chinese Academy of Sciences, Beijing 100049, PR China.
authors: Xianju
authors: X
authors: Liu
authors: Beijing Key Laboratory of Grape Science and Enology and Key Laboratory of Plant Resource, Institute of Botany, the Chinese Academy of Sciences, Beijing 100093, PR China.|University of Chinese Academy of Sciences, Beijing 100049, PR China.
authors: Zhan
authors: Z
authors: Zhang
authors: Beijing Key Laboratory of Grape Science and Enology and Key Laboratory of Plant Resource, Institute of Botany, the Chinese Academy of Sciences, Beijing 100093, PR China.|University of Chinese Academy of Sciences, Beijing 100049, PR China.
authors: Yi
authors: Y
authors: Wang
authors: Beijing Key Laboratory of Grape Science and Enology and Key Laboratory of Plant Resource, Institute of Botany, the Chinese Academy of Sciences, Beijing 100093, PR China.
authors: Wei
authors: W
authors: Duan
authors: Beijing Key Laboratory of Grape Science and Enology and Key Laboratory of Plant Resource, Institute of Botany, the Chinese Academy of Sciences, Beijing 100093, PR China.
authors: Shaohua
authors: S
authors: Li
authors: Beijing Key Laboratory of Grape Science and Enology and Key Laboratory of Plant Resource, Institute of Botany, the Chinese Academy of Sciences, Beijing 100093, PR China.|Sino-Africa Joint Research Center, Chinese Academy of Sciences, Wuhan 430074, PR China.
authors: Zhenchang
authors: Z
authors: Liang
mesh_terms: D001483:Base Sequence; D053898:Biosynthetic Pathways; D064113:CRISPR-Cas Systems; D004252:DNA Mutational Analysis; D024861:Genetic Enhancement; D018745:Genome, Plant; D054643:INDEL Mutation; D016296:Mutagenesis; D010088:Oxidoreductases / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics*; D013644:Tartrates / Q000378:metabolism; D027843:Vitis / Q000201:enzymology / Q000235:genetics*
chemical_list: D010940:Plant Proteins; D013644:Tartrates; D010088:Oxidoreductases; C029768:tartaric acid