Link to home

Editing citrus genome via SaCas9/sgRNA system

Hongge Jia: CREC, University of Florida

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