%0 Journal Article
%T A Simple PCR-based Strategy for the Introduction of Point Mutations in the
Yeast Saccharomyces cerevisiae via CRISPR/Cas9 | Insight Medical Publishing
%A Guohui Hu
%A Hai Rao
%A Haili Cheng
%A Shiwen Luo
%A Xin Gan
%J Journal Impact Factor: 1.48
%D 2018
%R 10.21767/2471-8084.100058
%X The methods currently employed for in vivo site-directed mutagenesis in yeast are laborious and/or inefficient. Recent developments of the CRISPR-based approaches hold great promise for genome editing, but its application in the yeast S. cerevisiae remains a time-consuming affair. The rate-limiting step in CRISPR-mediated genetic engineering in yeast is the incorporation of the guide sequences, which target Cas9 to relevant chromosomal locus, into the relevant yeast vectors. Here we present a PCR-based strategy to introduce specific point mutation into the yeast CDC48 gene via CRISPR. Our method eliminates the need for special dam- strain and markedly shortens the elaborate multi-step cloning process, leading to significant savings in time, labor and cost.
%U http://biochem-molbio.imedpub.com/a-simple-pcrbased-strategy-for-the-introduction-of-point-mutations-in-theyeast-saccharomyces-cerevisiae-via-crisprcas9.php?aid=22035