Genome editing: CRISPR-Cas9

Jakob B. Hoof, Christina S. Nødvig, Uffe H. Mortensen*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

Abstract

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

Keywords

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

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