RNA-Mediated Gene Silencing in Monokaryons and Dikaryons of Schizophyllum commune

• Published in: Applied and Environmental Microbiology Vol. 72; no. 2; pp. 1267 - 1269
• Main Authors: de Jong, Jan F, Deelstra, Heine J, Wösten, Han A. B, Lugones, Luis G
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.02.2006
• Subjects: Base Sequence; Basidiomycetes; Biological and medical sciences; cell biology; dikaryons; DNA, Fungal - genetics; Fundamental and applied biological sciences. Psychology; fungal proteins; Fungal Proteins - biosynthesis; Fungal Proteins - genetics; Fungi; gene silencing; gene transfer; genes; Genes, Fungal; genetic transformation; Genetics; hydrophobin; messenger RNA; Methods; Microbiology; monokaryons; mutants; mutation; Nucleic Acid Conformation; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Ribonucleic acid; RNA; RNA Interference; RNA, Fungal - chemistry; RNA, Fungal - genetics; RNA, Fungal - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; SC15 gene; Schizophyllum - genetics; Schizophyllum - metabolism; Schizophyllum commune; Fungi; Gene silencing; Basidiomycetes; Schizophyllum commune; RNA; Thallophyta
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/AEM.72.2.1267-1269.2006

Disruption of genes by homologous recombination occurs at a low frequency in the basidiomycete Schizophyllum commune. For instance, the SC3 and SC15 genes were inactivated at frequencies of 1 and 5%, respectively. As an alternative to disruption, we used gene silencing through the introduction of a hairpin construct. The SC15 gene, which encodes an abundantly secreted structural protein, was silenced at a frequency of 80% in monokaryons of S. commune after introduction of a hairpin construct of the gene. Silencing also occurred in dikaryons in which one of the partners was not a silenced strain. The silencing mechanism resembles RNAi in other filamentous fungi and is a powerful tool for the functional analysis of genes expressed in monokaryons or dikaryons.