# Genetic Manipulation in Sporothrix Species: Molecular Tools, Challenges, and Applications

## TL;DR

This paper reviews molecular tools for studying Sporothrix fungi, which cause sporotrichosis, and highlights recent advances in genetic manipulation techniques.

## Contribution

The paper provides a comprehensive overview of emerging genetic tools and their impact on understanding Sporothrix virulence.

## Key findings

- Recent genetic tools like CRISPR/Cas9 are enabling functional studies of Sporothrix virulence factors.
- Molecular techniques are improving understanding of gene function in clinical and environmental Sporothrix strains.
- Challenges remain in applying these tools broadly due to technical limitations in fungal transformation.

## Abstract

Sporothrix species are thermally dimorphic fungi responsible for sporotrichosis, a globally prevalent subcutaneous mycosis and an emerging zoonotic threat, particularly in South America. The high virulence of Sporothrix brasiliensis and its efficient transmission from cats to humans have intensified recent outbreaks, underscoring the importance of understanding the pathogenic mechanisms. While several putative virulence factors have been identified, such as melanin production, cell wall remodeling, extracellular vesicles, and thermotolerance, functional studies remain hampered by limited molecular tools. Recent advances, including random mutagenesis, protoplast-mediated transformation, Agrobacterium tumefaciens-mediated transformation, RNA interference and CRISPR/Cas9-based genome editing, are changing this landscape. These methods have enabled the functional validation of key virulence factors and the investigation of gene function in both environmental and clinical strains. In this review, we summarize the genetic toolbox available for Sporothrix, outline current challenges, and discuss how these strategies are reshaping the study of fungal virulence and host–pathogen interactions.