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.
CITATION STYLE
Yoshinaga, Y., Daum, C., Guifen, H., & O´Malley, R. (2018). Chapter 11 Genome Sequencing. Fungal Genomics: Methods and Protocols, 1775, 37–52.
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