A user-friendly and streamlined protocol for CRISPR/Cas9 genome editing in budding yeast
Daniele Novarina, Ανδριάνα Κουτσούμπα, Andreas Milias‐Argeitis
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).
Topics & Concepts
CRISPRGenome editingCas9Computer scienceProtocol (science)Computational biologyGenomeSubgenomic mRNAPlasmidBiologyGeneticsDNAGeneAlternative medicineMedicinePathologyCRISPR and Genetic EngineeringInsect symbiosis and bacterial influencesGenetics, Aging, and Longevity in Model Organisms