利用CRISPR-Cas9基因编辑技术对CSSL37的YSA基因进行定点编辑

Editing of YSA Gene of CSSL37 Based on CRISPR-Cas9 Gene Editing System

水稻是中国乃至世界重要的粮食作物之一,作物染色体片段置换系群体是作物新资源的挖掘、数量遗传研究和杂种优势利用研究的理想工具和材料。CRISPR-Cas9技术是近年发展起来的一种快速基因编辑技术,通过该技术已对多种植物的基因组进行了编辑。YSA的突变体ysa,由于在编码区5个碱基的缺失导致白化的性状,ysa在3叶期以前发育出白化叶,之后叶片逐渐变绿,到6叶期完全恢复成正常绿色。本研究利用CRISPR-Cas9技术,构建同时含有2个sg RNA载体,以CSSL37株系为材料,通过对YSA基因进行定向编辑和修饰。结果表明,获得CSSL37株系的转基因苗20株,有18个株系在目标区域内序列均发生变化,其中有2个株系在T0代已经纯合,出现目标性状的表型。该研究为进一步利用基因编辑体系和创制优异的水稻新材料提供了帮助。

Rice is one of the important food crops in China and even in the world. Chromosome segment substitution line are the most ideal tools and materials for genetic research like excavating new resources, quantitative genetic studies and the utilization of heterosis. CRISPR-Cas9-based gene editing technology has developed rapidly in recent years. The genome of many plants has been modified by this biotechnology. Ysa, the mutant of YSA, which showed a 5-bp deletion in ORF4, led to the leaves ofysa appeared albinic albino before the three-leaf stage, but gradually tumed green from the four-leaf stage onward and appeared normal green after the six-leaf stage. In this study, we took CSSL37 as the material and used CRISPR-Cas9 technology to construct two sgRNA vectors simultaneously, which edited and modified YSA genes directly. The research results showed that there were 18 strains within the target area had sequence change in 20 plants, and 2 strains in To was homozygous, which showed phenotype of the target traits. The study will be helpful for further utilizing the genetic editing system and creating the excellent new materials for rice.