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

<i>Sporothrix</i> 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 <i>Sporothrix brasiliensis</i> 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, <i>Agrobacterium tumefaciens</i>-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 <i>Sporothrix</i>, outline current challenges, and discuss how these strategies are reshaping the study of fungal virulence and host-pathogen interactions.