Single and Multiplexed Gene Editing in Ustilago maydis Using CRISPR-Cas9

Mariana Schuster; Christine Trippel; Petra Happel; Daniel Lanver; Stefanie Reißmann; Regine Kahmann

doi:10.21769/BioProtoc.2928

Single and Multiplexed Gene Editing in Ustilago maydis Using CRISPR-Cas9

Bio Protoc. 2018 Jul 20;8(14):e2928. doi: 10.21769/BioProtoc.2928.

Authors

Mariana Schuster¹, Christine Trippel¹, Petra Happel¹, Daniel Lanver¹, Stefanie Reißmann¹, Regine Kahmann¹

Affiliation

¹ Max Planck Institute for Terrestrial Microbiology, Department of Organismic Interactions, Marburg, Germany.

Abstract

The smut fungus Ustilago maydis is an established model organism for elucidating how biotrophic pathogens colonize plants and how gene families contribute to virulence. Here we describe a step by step protocol for the generation of CRISPR plasmids for single and multiplexed gene editing in U. maydis. Furthermore, we describe the necessary steps required for generating edited clonal populations, losing the Cas9 containing plasmid, and for selecting the desired clones.

Keywords: CRISPR-Cas9; Gene editing; Multiplexing; Smut fungi; Ustilago maydis.