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

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).

SUBMITTER: Novarina D

PROVIDER: S-EPMC9192979 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

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

Novarina Daniele D Koutsoumpa Andriana A Milias-Argeitis Andreas A

STAR protocols 20220610 2

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 an ...[more]

PMID: 35712010

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