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

Carlos Lax, María Isabel Navarro-Mendoza, Carlos Pérez‐Arques, Eusebio Navarro, Francisco E. Nicolás, Victoriano Garre

2022STAR Protocols13 citationsDOIOpen Access PDF

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).

Topics & Concepts

CRISPRGenome editingHomologous recombinationComputational biologyBiologyCas9Transformation (genetics)GeneticsGenomeDNAGeneAntifungal resistance and susceptibilityFungal Infections and StudiesPlant-Microbe Interactions and Immunity
Transformation and CRISPR-Cas9-mediated homologous recombination in the fungus Rhizopus microsporus | Litcius