RNA interference in the pathogenic fungus Cryptococcus neoformans.
Cryptococcus neoformans is a pathogenic fungus responsible for serious disease in immunocompromised individuals. This organism has recently been developed as an experimental system, with initiation of a genome project among other molecular advances. However, investigations of Cryptococcus are hampered by the technical difficulty of specific gene replacements. RNA interference, a process in which the presence of double-stranded RNA homologous to a gene of interest results in specific degradation of the corresponding message, may help solve this problem. We have shown that expression of double-stranded RNA corresponding to portions of the cryptococcal CAP59 and ADE2 genes results in reduced mRNA levels for those genes, with phenotypic consequences similar to that of gene disruption. The two genes could also be subjected to simultaneous interference through expression of chimeric double-stranded RNA. Specific modulation of protein expression through introduction of double-stranded RNA thus operates in C. neoformans, which is the first demonstration of this technique in a fungal organism. Use of RNA interference in Cryptococcus should allow manipulation of mRNA levels for functional analysis of genes of interest and enable efficient exploration of genes discovered by genome sequencing.
Liu
Hong
H
Department of Molecular Microbiology, Washington University School of Medicine, Saint Louis, Missouri 63110, USA.
Cottrell
Tricia R
TR
Pierini
Lynda M
LM
Goldman
William E
WE
Doering
Tamara L
TL
D002262:Carboxy-Lyases / Q000235:genetics / Q000502:physiology; D003455:Cryptococcus neoformans / Q000235:genetics* / Q000502:physiology; D015536:Down-Regulation / Q000502:physiology; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology; D015966:Gene Expression Regulation, Fungal; D012331:RNA, Fungal / Q000235:genetics* / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics* / Q000502:physiology
C087781:CAP59 protein, Cryptococcus neoformans; D005656:Fungal Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D002262:Carboxy-Lyases; C022378:phosphoribosylaminoimidazole carboxylase