Gene Silencing by RNA Interference in the Koji Mold<i>Aspergillus oryzae</i>

Ikeda, Ryoko; Ohkita, Yuka; Hayashi, Risa; Sakamoto, Kazutoshi; Akita, Osamu

doi:10.1271/bbb.60405

Cited by 36 publications

(27 citation statements)

References 33 publications

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“…Accordingly, both Tet-On modules allow gene silencing, which is essential for studying the cellular function of any gene of interest by conditional promoter replacement. This screening approach has proven extremely useful for the identification of virulence determinants and drug targets in fungal pathogens (1,13) and represents a superior alternative to the commonly used gene deletion strategy or the recent RNA interference (RNAi)-mediated gene silencing approach (14,15). Of special interest is the fact that both Tet-On constructs differ in their response toward the endogenous inducer doxycycline, with the p oliC version resulting in apparently sufficient expression of the rho1 gene product at lower concentrations at the costs of an apparently higher basal expression.…”

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confidence: 99%

Upgrading Fungal Gene Expression on Demand: Improved Systems for Doxycycline-Dependent Silencing in Aspergillus fumigatus

Helmschrott

Sasse

Samantaray

et al. 2013

Appl Environ Microbiol

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confidence: 99%

Upgrading Fungal Gene Expression on Demand: Improved Systems for Doxycycline-Dependent Silencing in Aspergillus fumigatus

Helmschrott

Sasse

Samantaray

et al. 2013

Appl Environ Microbiol

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“…This dsRNA is processed into 21-to 25-nucleotide fragments which associate with a nuclease complex (RNA-induced silencing complex) and are used as a guide for homology-dependent degradation of the target mRNA (11,14). RNAi has been used as a method to study gene function and for specific inhibition of gene expression in a range of organisms, including several basidiomycetes and ascomycetes such as Aspergillus fumigatus (30), As-pergillus oryzae (45), Coprinus cinereus (32,44), Schizophyllum commune (12), Cryptococcus neoformans (27), and Neurospora crassa (10). This procedure may be particularly useful for the simultaneous suppression of closely related genes, as well as the partial suppression of essential genes (31,36).…”

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confidence: 99%

Gene Silencing by RNA Interference in the White Rot Fungus Phanerochaete chrysosporium

Matityahu

Hadar

Dosoretz

et al. 2008

Appl Environ Microbiol

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The effectiveness of RNA interference (RNAi) is demonstrated in the lignin-degrading fungus Phanerochaete chrysosporium. The manganese-containing superoxide dismutase gene (MnSOD1) was used as the target for RNAi. The plasmid constructed for gene silencing contained a transcriptional unit for hairpin RNA expression. Significantly lower MnSOD expression at both the mRNA and protein activity levels was detected in RNAi transformants. Furthermore, even though P. chrysosporium possesses three copies of the MnSOD gene, this RNAi construct was sufficient to decrease the enzymatic activity by as much as 70% relative to control levels. Implementation of the RNAi technique in P. chrysosporium provides an alternative genetic tool for studies of gene function, particularly of essential genes or gene families.

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“…On the one hand, rearrangements may affect copy number determinations (even using Southern blotting techniques, which frequently miss smaller variations), and on the other hand, agarose gel analysis of amplified segments of genomic DNA is insufficient to provide reliable quantification. Nevertheless, combined analysis of these two methods may be used: Fr example, copy numbers of pXLNRir do not correlate with a higher degree of silencing efficiency, a situation similar to others reported (Nakayashiki et al 2005;Nicolas et al 2003;Tanguay et al 2006;Fitzgerald et al 2004;Henry et al 2007;Yamada et al 2007;Walti et al 2006). Apart from genome instability and the occurrence of rearrangements, other factors may be responsible for a lower silencing efficiency of integrated hairpin-expression units.…”

Section: Discussionmentioning

confidence: 67%

Efficient cloning system for construction of gene silencing vectors in Aspergillus niger

Oliveira

Veen

Graaff

2008

Appl Microbiol Biotechnol

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An approach based on Gateway recombination technology to efficiently construct silencing vectors was developed for use in the biotechnologically important fungus Aspergillus niger. The transcription activator of xylanolytic and cellulolytic genes XlnR of A. niger was chosen as target for gene silencing. Silencing was based on the expression vector pXLNRir that was constructed and used in co-transformation. From all the strains isolated (N=77), nine showed poor xylan-degrading activities in two semiquantitative plate assays testing different activities for xylan degradation. Upon induction on D-xylose, transcript levels of xlnR were decreased in the xlnR-silenced strains, compared to a wild-type background. Under these conditions, the transcript levels of xyrA and xynB (two genes regulated by XlnR) were also decreased for these xlnRsilenced strains. These results indicate that the newly developed system for rapid generation of silencing vectors is an effective tool for A. niger, and this can be used to generate strains with a tailored spectrum of enzyme activities or product formation by silencing specific genes encoding, e.g., regulators such as XlnR.

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Gene Silencing by RNA Interference in the Koji MoldAspergillus oryzae

Cited by 36 publications

References 33 publications

Upgrading Fungal Gene Expression on Demand: Improved Systems for Doxycycline-Dependent Silencing in Aspergillus fumigatus

Upgrading Fungal Gene Expression on Demand: Improved Systems for Doxycycline-Dependent Silencing in Aspergillus fumigatus

Gene Silencing by RNA Interference in the White Rot Fungus Phanerochaete chrysosporium

Efficient cloning system for construction of gene silencing vectors in Aspergillus niger

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