Version 2.0 of the Hermes transposon-based mutagenesis system for prototrophic mutant pool generation in Candida glabrata.

Although the original Hermes transposon mutagenesis system has proven successful for saturated insertional mutagenesis in Candida glabrata, its reliance on uracil auxotrophy may limit its applicability under nutrient-restricted host environments. To overcome this limitation, Zaeske et al. (Microbiol Spectr 14:e0084826, 2026, https://doi.org/10.1128/spectrum.00848-26) recently developed a prototrophic alternative that uses cycloheximide (CHX) instead of 5-fluoroorotic acid (5-FOA) to enrich for transposon insertion mutants, generating high-complexity pools. This system provides a valuable resource for investigating the genetic and cellular determinants of colonization and pathogenicity in this important fungal pathogen.