Near 100% efficient homology-dependent genome engineering in the human fungal pathogen Cryptococcus neoformans.

ABSTRACT: We recently described CRISPR/Cas9-based short homology-dependent genome engineering in the human fungal pathogen Cryptococcus neoformans, a haploid budding yeast that is the most common cause of fungal meningitis and an emerging model organism. This was achieved by electroporation of strains stably expressing a codon-optimized Cas9 with 2 separate DNA molecules, one encoding a selectable marker flanked by short homology arms and a second encoding a sgRNA under the control of the U6 snRNA promoter. However, the efficiency of desired homology-dependent repair relative to undesired non-homologous end-joining (NHEJ) events can be low and variable. Here, we describe methods and strains enabling extremely efficient (∼99%) homology-dependent genome editing in C. neoformans. This high-efficiency m

SUBMITTER: Nalley MJ 

PROVIDER: S-EPMC12341884 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature