Transformation and CRISPR-Cas9-mediated homologous recombination in the fungus Rhizopus microsporus

Carlos Lax; María Isabel Navarro-Mendoza; Carlos Pérez-Arques; Eusebio Navarro; Francisco Esteban Nicolás; Victoriano Garre

doi:10.1016/j.xpro.2022.101237

Transformation and CRISPR-Cas9-mediated homologous recombination in the fungus Rhizopus microsporus

STAR Protoc. 2022 Mar 14;3(1):101237. doi: 10.1016/j.xpro.2022.101237. eCollection 2022 Mar 18.

Authors

Carlos Lax¹, María Isabel Navarro-Mendoza², Carlos Pérez-Arques², Eusebio Navarro¹, Francisco Esteban Nicolás¹, Victoriano Garre¹

Affiliations

¹ Departamento de Genética y Microbiología, Facultad de Biología, Universidad de Murcia, 30100 Murcia, Spain.
² Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.

Abstract

Here, we describe a reliable approach for targeted DNA integrations in the genome of R. microsporus, one of the main causal agents of mucormycosis. We provide a strategy for stable, targeted integration of DNA templates by homologous recombination (HR) based on the CRISPR-Cas9 technology. This strategy opens a wide range of possibilities for the genetic modification of R. microsporus and will be useful for the study of mucormycosis. For complete details on the use and execution of this protocol, please refer to Lax et al. (2021).

Keywords: Biotechnology and bioengineering; Genetics; Microbiology; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems* / genetics
DNA
Homologous Recombination
Mucormycosis* / genetics
Rhizopus / genetics

Substances

DNA

Supplementary concepts

Rhizopus microsporus