Genome editing: CRISPR-Cas9

Research output: Chapter in Book/Report/Conference proceedingBook chapter – Annual report year: 2018Researchpeer-review

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In the present chapter, we present the protocols and guidelines to facilitate implementation of CRISPR-Cas9 technology in fungi where few or no genetic tools are in place. Hence, we firstly explain how to identify dominant markers for genetic transformation. Secondly, we provide a guide for construction of Cas9/sgRNA episomal expression vectors. Thirdly, we present how to mutagenize reporter genes to explore the efficiency of CRISPR-Cas9 in the relevant fungus and to ease subsequent CRISPR-mediated genetic engineering. Lastly, we describe how to make CRISPR-mediated marker-dependent and marker-free gene targeting.

Original languageEnglish
Title of host publicationFungal Genomics: methods and protocols
EditorsRonald P. de Vries, Adrian Tsang, Igor V. Grigoriev
Volume1775
PublisherHumana Press
Publication date2018
Edition2
Pages119-132
Chapter11
ISBN (Print)978-1-4939-7803-8
ISBN (Electronic)978-1-4939-7804-5
DOIs
Publication statusPublished - 2018
SeriesMethods in Molecular Biology
Volume1775
ISSN1064-3745
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • CRISPR, Gene editing, Gene targeting, Mutagenesis, Nonmodel fungi

ID: 153819323