title: Gene silencing by RNA interference in the koji mold Aspergillus oryzae.
abstract: We found the orthologous genes required for RNA interference (RNAi) in the Aspergillus oryzae genome database, and constructed a set of tools for gene silencing using RNAi in A. oryzae. This system utilizes compatible restriction enzyme sites so that only a single target gene fragment is required to create the hairpin RNA cassette. For ease of handling, we also separated the construction of the hairpin RNA cassette for the target gene from its subsequent introduction into the expression vector. Using the brlA gene as a target for RNAi, we detected decreased mRNA levels and a delayed conidiation phenotype in the transformants. Furthermore, even though A. oryzae possesses three copies of the alpha-amylase gene, a single copy of an alpha-amylase RNAi construct was sufficient to downregulate the mRNA levels and decrease the enzymatic activity to 10% of control levels. Gene silencing by RNAi should provide a powerful genetic tool for post-genomic studies of the industrially important fungus A. oryzae.
authors.affiliation: National Research Institute of Brewing, Japan. yamada@nrib.go.jp
authors.forename: Osamu
authors.initials: O
authors.lastname: Yamada
authors.forename: Ryoko
authors.initials: R
authors.lastname: Ikeda
authors.forename: Yuka
authors.initials: Y
authors.lastname: Ohkita
authors.forename: Risa
authors.initials: R
authors.lastname: Hayashi
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Sakamoto
authors.forename: Osamu
authors.initials: O
authors.lastname: Akita
mesh_terms: D001236:Aspergillus oryzae / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D016681:Genome, Fungal / Q000235:genetics; D034622:RNA Interference; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism
chemical_list: D014157:Transcription Factors; D000516:alpha-Amylases