Cas9-mediated gene editing in Saccharomyces cerevisiae

Abstract

We describe an easy, e cient, and inexpensive way to clone gRNAs into plasmid vectors carrying Cas9.The method involves designing two 25 nt complementary sequences that can be duplexed and subsequently ligated into the BplI restriction site of the plasmid vector.Using the above method, we have been able to introduce point mutations and deletions in varying sizes by using 80-nt single-stranded DNA, PCR products or gBlocks as templates.