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

ABSTRACT 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.