A user-friendly and streamlined protocol for CRISPR/Cas9 genome editing in budding yeast

  • STAR Protoc. 2022 Jun 10;3(2):101358. doi: 10.1016/j.xpro.2022.101358.
Daniele Novarina  1 ,  Andriana Koutsoumpa  1 ,  Andreas Milias-Argeitis  1
Affiliations
  • 1. Molecular Systems Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, the Netherlands.
PMID: 35712010 DOI: 10.1016/j.xpro.2022.101358
Abstract

CRISPR/Cas9 technology allows accurate, marker-less genome editing. We report a detailed, robust, and streamlined protocol for CRISPR/Cas9 genome editing in Saccharomyces cerevisiae, based on the widely used MoClo-Yeast Toolkit (https://www.addgene.org/kits/moclo-ytk/). This step-by-step protocol guides the reader from sgRNA design to verification of the desired genome editing event and provides preassembled plasmids for cloning the sgRNA(s), making this technology easily accessible to any yeast research group. For complete details on the use and execution of this protocol, please refer to Novarina et al. (2021).

Keywords
CRISPR; Genetics; Model Organisms; Molecular Biology.