Comparison of CRISRP-Cas9 system and mazF-mediated method for large deletions in Saccharomyces cerevisiae

Abstract:

[Objective] The objective of this research was to compare CRISRP-Cas9 system and mazF-mediated method for large deletions in Saccharomyces cerevisiae.[Methods] We made a 26.5 kb deletion from YKL072W to YKL061W using the foresaid two methods. The two methods were analyzed from the perspective of transformation rate and accuracy of deletion.[Results] There were five colonies appeared on the plate in average using CRISRP-Cas9 system and all of these were correct. And 100 colonies were observed using the mazF-mediated method and the accuracy was 93%, a little bit lower than CRISPR-Cas9 system.[Conclusion] Both methods are good to make large deletion in Saccharomyces cerevisiae. The CRISRP-Cas9 system has a high accuracy and easy to use, the mazF-mediated method is stable and does not need the PAM sequence.