AUTHOR=Jia Hongge , Xu Jin , Orbović Vladimir , Zhang Yunzeng , Wang Nian 

TITLE=Editing Citrus Genome via SaCas9/sgRNA System

JOURNAL=Frontiers in Plant Science

VOLUME=8

YEAR=2017

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2017.02135

DOI=10.3389/fpls.2017.02135

ISSN=1664-462X

ABSTRACT=<p>SaCas9/sgRNA, derived from <italic>Staphylococcus aureus</italic>, is an alternative system for genome editing to <italic>Streptococcus pyogenes</italic> 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 <italic>Arabidopsis</italic>, 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 <italic>CsPDS</italic> and <italic>Cs2g12470</italic>. Subsequently, binary vector GFP-p1380N-SaCas9/35S-sgRNA1:AtU6-sgRNA2 was developed to edit two target sites of <italic>Cs7g03360</italic> 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.</p>