Editing Citrus Genome via SaCas9/sgRNA System

• DOI: 10.3389/fpls.2017.02135
• Publication Date: 2017-12-12T04:40:01Z
• Authors: Hongge Jia; Jin Xu; Vladimir Orbović; Yunzeng Zhang; Nian Wang
• Keywords: Subgenomic mRNA

SaCas9/sgRNA, derived from Staphylococcus aureus, is an alternative system for genome editing to Streptococcus pyogenes SpCas9/sgRNA. The smaller SaCas9 recognizes a different protospacer adjacent motif (PAM) sequence SpCas9. SaCas9/sgRNA has been employed edit the genomes of Arabidopsis, tobacco and rice. In this study, we aimed test its potential in citrus. Transient expression Duncan grapefruit via Xcc-facilitated agroinfiltration showed it can successfully modify CsPDS Cs2g12470. Subsequently, binary vector GFP-p1380N-SaCas9/35S-sgRNA1:AtU6-sgRNA2 was developed two target sites Cs7g03360 transgenic Carrizo citrange. Twelve GFP-positive transformants were established, designated as #Cz1 #Cz12. Based on targeted next generation sequencing results, mutation rates targets ranged 15.55 39.13% sgRNA1 49.01 79.67% sgRNA2. Therefore, be used tool SpCas9/sgRNA citrus editing.