SCR7和RS-1对酿酒酵母中CRISPR/Cas9编辑效率的影响
Effects of SCR7 and RS-1 on the Editing Efficiency of CRISPR/Cas9 in Saccharomyces Cerevisiae
倪江萍 1杨兰 1张轩2
作者信息
- 1. 宁夏医科大学基础医学院,银川 750004
- 2. 宁夏回族自治区人民医院宁夏医科大学第三临床医学院,银川 750002
- 折叠
摘要
目的 研究SCR7 和RS-1 对酿酒酵母中CRISPR/Cas9 编辑效率的影响.方法 CRISPR/Cas9 系统能特异性地切断DNA双链,产生双链断裂(DSB),在真核细胞中DSB的修复方式主要有两种,非同源末端连接(NHEJ)和同源重组修复(HDR).小分子化合物SCR7 和RS-1 在DNA修复过程中能通过抑制NHEJ或增强HDR提高 CRISPR/Cas9 精准基因编辑效率.本研究将探究用不同浓度的SCR7 和RS-1 分别处理酿酒酵母,验证YPL062W 基因的删除效率.结果 本研究发现,添加 5、10、15、20 μmol SCR7 后,YPL062W 基因的删除效率分别为 80%、90%、90%和 80%,添加 1、5、10、15、20 μmol RS-1 后,YPL062W 基因的删除效率分别为70%、70%、80%、70%和 70%,添加小分子化合物组合 10 μmol SCR7+10 μmol RS-1 和 15 μmol SCR7+10 μmol RS-1 后,YPL062W 基因的删除效率分别为 70%和 80%.结论 SCR7 和 RS-1 能显著增强酿酒酵母中CRISPR/Cas9 系统基因编辑效率.
Abstract
Objective To investigate the effects of SCR7 and RS-1 on the editing efficiency of CRISPR/Cas9 in saccharomyces cerevisiae.Methods The CRISPR/Cas9 system could specifically cleave DNA double strands,producing double-strand break(DSB).In eukaryotic cells,there were two main repair methods for DSB:non-homologous end joining(NHEJ)and homology directed repair(HDR).Small molecule compounds SCR7 and RS-1 could improve the precision gene editing efficiency of CRISPR/Cas9 during DNA repair by inhibiting NHEJ or enhancing HDR.The deletion efficiency of the reporter gene YPL062W was verified by treating saccharomyces cerevisiae with different concentrations of SCR7 and RS-1.Results Research found that the deletion efficiency of the YPL062W gene was 70%,90%,90%and 80%,respectively after adding 5,10,15 and 20 μmol SCR7.Adding 1,5,10,15,20 μmol RS-1,the deletion efficiency of the YPL062W gene was 70%,70%,80%,70%and 70%,respectively.Adding small molecule compound combinations 10 μmol SCR7+10 μmol RS-1 and 15 μmol SCR7+10 μmol RS-1,the deletion efficiencies of the YPL062W gene were 70%and 80%respectively.Conclusion SCR7 and RS-1 can significantly enhance the gene editing efficiency of the CRISPR/Cas9 system in saccharomyces cerevisiae.
关键词
CRISPR/Cas9系统/SCR7/RS-1/酿酒酵母Key words
CRISPR/Cas9 system/SCR7/RS-1/saccharomyces cerevisiae引用本文复制引用
基金项目
宁夏医科大学校级项目(XM2020013)
出版年
2024
NETL
NSTL
万方数据