Antisense repression in Cryptococcus neoformans as a laboratory tool and potential antifungal strategy.

Published

Journal Article

Antisense repression was used as a method to alter gene function in the human-pathogenic fungus Cryptococcus neoformans. The calcineurin A gene (CNA1) and the laccase gene (LAC1) were targeted since disruption of these loci results in phenotypes that are easy to screen (temperature sensitivity and lack of melanin, respectively). Serotype D yeasts were transformed with a plasmid containing the CNA1 cDNA in an antisense orientation under the control of the inducible GAL7 promoter, and serotype A yeasts were transformed with a plasmid containing the LAC1 cDNA in an antisense orientation under the control of the constitutive actin promoter. The calcineurin transformants demonstrated a temperature-sensitive phenotype only when grown on galactose, and the laccase transformants had decreased melanin production. Northern blot analysis of the calcineurin antisense transformants confirmed that the inducible phenotype was associated with a decrease in the native CNA1 transcript levels. Furthermore, it was possible to modestly impair growth of C. neoformans at 37 degrees C by using a 30 bp antisense oligonucleotide targeting CNA1. Antisense repression is now available as a tool for molecular studies in this organism, and may be applicable to other human-pathogenic fungi that have less amenable genetic systems.

Full Text

Duke Authors

Cited Authors

  • Gorlach, JM; McDade, HC; Perfect, JR; Cox, GM

Published Date

  • January 2002

Published In

Volume / Issue

  • 148 / Pt 1

Start / End Page

  • 213 - 219

PubMed ID

  • 11782513

Pubmed Central ID

  • 11782513

International Standard Serial Number (ISSN)

  • 1350-0872

Digital Object Identifier (DOI)

  • 10.1099/00221287-148-1-213

Language

  • eng

Conference Location

  • England