水稻OsDTH14基因编辑突变体的创建与功能初探

he CRISPR/Cas9 gene editing system is a genome editing tool based on the bacterial immune system, which has been widely applied in the field of plants in recent years. It uses single guide RNA (sgRNA) to guide Cas9 nuclease to recognize and cleave target DNA, causing double strand breaks, and then creating gene knockout, insertion, or replacement through DNA repair mechanisms. In this study, CRISPR/Cas9 technology was adopted to design two targeting sites at the first exon of the OsDTH14 gene in rice, and a CRISPR/Cas9 gene editing vector was constructed. Then using Agrobacterium-mediated rice genetic transformation method, the vector was introduced into theGeneration of mutants of OsDTH14 using CRISPR/Cas9 system and preliminary study on the function in rice rice variety Zhonghua 11 (Zh11) to obtain OsDTH14-CRISPR/Cas9 gene edited mutant plants. A homozygous editing mutant and a heterozygous mutant were obtained and validated by sequencing analysis and PCR. This work laid the foundation for further detailed research on the function of OsDTH14.