Reach Us +44-1904-929220

Abstract

A Simple PCR-based Strategy for the Introduction of Point Mutations in the Yeast Saccharomyces cerevisiae via CRISPR/Cas9

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.


Author(s):

Guohui Hu, Shiwen Luo, Hai Rao, Haili Cheng and Xin Gan



Abstract | Full-Text | PDF

Share this  Facebook  Twitter  LinkedIn  Google+
Flyer image

Abstracted/Indexed in

  • Index Copernicus
  • Google Scholar
  • Directory of Research Journal Indexing (DRJI)
  • WorldCat
  • Publons
  • Scholarsteer