生物技术通报2025，Vol.41Issue(3)：62-70,9.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0784
优化sgRNA提高塔宾曲霉基因编辑效率
Improving the Efficiency of Gene Editing by Optimizing sgRNA in Aspergillus tubingensis
摘要
Abstract
[Objective]In the CRISPR/Cas9 gene editing system,sgRNA is one of the important gene editing elements.It binds to the Cas9 protein and complements the genomic DNA through the spacer sequence to guide the Cas9 protein to accurately cut the genome.In order to improve the gene editing efficiency of Aspergillus tubingensis,sgRNA optimization is a feasible strategy.[Method]The promoter and hairpin structure of sgRNA were optimized,and the gene editing efficiency was verified in A.tubingensis.[Result]The gene editing efficiency of sgRNA with"lock"structure was 9.37%higher than that of the control group when gene editing was performed by RNP method.When sgRNA was expressed in vivo,the gene editing efficiency of tRNAGly15 and tRNAGly17 promoters was 14%-16%higher than that of 5S rRNA promoter,respectively.Using the tRNAGly15 promoter to express sgRNA with a"lock"structure increased the white spore rate and increased the gene editing efficiency of A.tubingensis to 96%.[Conclusion]The"lock"structure and two promotors of tRNAGly15 and tRNAGly17 may improve the gene editing efficiency of A.tubingensis.关键词
塔宾曲霉/基因编辑效率/启动子/"lock"结构Key words
Aspergillus tubingensis/gene editing efficiency/promoter/"lock"structure引用本文复制引用
梁丽存,王克芬,宋祖洹,刘梦婷,李佳玉,罗会颖,姚斌,杨浩萌..优化sgRNA提高塔宾曲霉基因编辑效率[J].生物技术通报,2025,41(3):62-70,9.基金项目
国家重点研发计划(2021YFC2100204),国家现代农业产业技术体系专项(CARS-41) （2021YFC2100204）
