Simple growth conditions improve targeted gene deletion in <i>Cryptococcus neoformans</i>.

ABSTRACT: Cryptococcus neoformans infections are a significant cause of morbidity and mortality among AIDS patients and the third most common invasive fungal infection in organ transplant recipients. The cryptococcal cell wall is very dynamic and can be modulated depending on growth conditions. It was reported that when C. neoformans is grown in unbuffered yeast nitrogen base (YNB) for 48 hours, the pH of the media drastically drops, and the cells start to shed their cell walls. With this observation, we sought to determine if YNB-grown cells could be used directly for genetic transformation. To test this, we targeted ADE2 using TRACE (transient CRISPR-Cas9 coupled with electroporation) in YNB-grown or competent cells. Deletion of the ADE2 gene results in red-pigmented co

SUBMITTER: Watson RG 

PROVIDER: S-EPMC12039239 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature