DNA-free genome editing in grapevine using CRISPR/Cas9 ribonucleoprotein complexes followed by protoplast regeneration
作者机构：Department of BiotechnologyUniversity of Verona37134 VeronaItaly
出 版 物：《Horticulture Research》 (园艺研究（英文）)
年 卷 期：2023年第10卷第1期
页 面：183-192页
核心收录：
学科分类：09[农学] 0902[农学-园艺学] 090201[农学-果树学]
基 金：This research was funded by the European Union’s Horizon 2020 research and innovation programme under Marie Skłodowska-Curie grant no.754345 awarded to SN the University of Verona in the framework of the Grant Ricerca di Base“Definition of master regulator genes of fruit ripening in grapevine”awarded to SZ and the Ministero delle Politiche Agricole Alimentari e Forestali(Mipaaf)in the framework of the BIOTECH-VITECH(CIG:8704614AB4)project awarded to SZ
摘 要：CRISPR/Cas9 genome editing technology can overcome many limitations of traditional breeding,offering enormous potential for crop improvement and food *** the direct delivery of Cas9-single guide RNA(sgRNA)ribonucleoprotein(RNP)complexes to grapevine(Vitis vinifera)protoplasts has been shown before,the regeneration of edited protoplasts into whole plants has not been ***,we describe an efficient approach to obtain transgene-free edited grapevine plants by the transfection and subsequent regeneration of protoplasts isolated from embryogenic *** proof of concept,a single-copy green f luorescent protein reporter gene(GFP)in the grapevine cultivar Thompson Seedless was targeted and knocked out by the direct delivery of RNPs to ***/Cas9 activity,guided by two independent sgRNAs,was confirmed by the loss of GFP f *** regeneration of GFP−protoplasts into whole plants was monitored throughout development,confirming that the edited grapevine plants were comparable in morphology and growth habit to wild-type *** report the first highly efficient protocol for DNA-free genome editing in grapevine by the direct delivery of preassembled Cas9-sgRNA RNP complexes into protoplasts,helping to address the regulatory concerns related to genetically modified *** technology could encourage the application of genome editing for the genetic improvement of grapevine and other woody crop plants.
维普期刊数据库