Effects of SCR7 and RS-1 on the Editing Efficiency of CRISPR/Cas9 in Saccharomyces Cerevisiae
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.
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