ArticleApplied and environmental microbiology2007
An efficient fungal RNA-silencing system using the DsRed reporter gene.
Article in Applied and environmental microbiology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 59 citations in OpenAlex.
- Review
- Cephalosporin C biosynthesis and fermentation in Acremonium chrysogenum.Applied microbiology and biotechnology · 2022Review
- Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi.AMB Express · 2020Article
- The construction and use of versatile binary vectors carrying pyrG auxotrophic marker and fluorescent reporter genes for Agrobacterium-mediated transformation of Aspergillus oryzae.World journal of microbiology & biotechnology · 2016Article
- Study on genetic engineering ofSynthetic and systems biotechnology · 2016Review
- A red fluorescent protein (DsRED) from Discosoma sp. as a reporter for gene expression in walnut somatic embryos.Plant cell reports · 2015Article
- Establishment of an efficient RNA silencing system in Trichoderma koningii using DsRed as a reporter.Folia microbiologica · 2013Article
- Infection structure-specific expression of β-1,3-glucan synthase is essential for pathogenicity of Colletotrichum graminicola and evasion of β-glucan-triggered immunity in maize.The Plant cell · 2013Article
- Development of an optimized tetracycline-inducible expression system to increase the accumulation of interleukin-10 in tobacco BY-2 suspension cells.BMC biotechnology · 2012Article
- The development and application of a multiple gene co-silencing system using endogenous URA3 as a reporter gene in Ganoderma lucidum.PloS one · 2012Article
- The phocein homologue SmMOB3 is essential for vegetative cell fusion and sexual development in the filamentous ascomycete Sordaria macrospora.Current genetics · 2011Article
- pHg/pSILBAγ vector system for efficient gene silencing in homobasidiomycetes: optimization of ihpRNA - triggering in the mycorrhizal fungus Laccaria bicolor.Microbial biotechnology · 2010Article
- Article
- Asexual cephalosporin C producer Acremonium chrysogenum carries a functional mating type locus.Applied and environmental microbiology · 2008Article
- Gene silencing by RNA Interference in the white rot fungus Phanerochaete chrysosporium.Applied and environmental microbiology · 2008Article
- Identification of a minimal cre1 promoter sequence promoting glucose-dependent gene expression in the beta-lactam producer Acremonium chrysogenum.Current genetics · 2008Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In filamentous fungi, RNA silencing is an attractive alternative to disruption experiments for the functional analysis of genes. We adapted the gene encoding the autofluorescent DsRed protein as a reporter to monitor the silencing process in fungal transformants. Using the cephalosporin C producer Acremonium chrysogenum, strains showing a high level of expression of the DsRed gene were constructed, resulting in red fungal colonies. Transfer of a hairpin-expressing vector carrying fragments of the DsRed gene allowed efficient silencing of DsRed expression. Monitoring of this process by Northern hybridization, real-time PCR quantification, and spectrofluorometric measurement of the DsRed protein confirmed that downregulation of gene expression can be observed at different expression levels. The usefulness of the DsRed silencing system was demonstrated by investigating cosilencing of DsRed together with pcbC, encoding the isopenicillin N synthase, an enzyme involved in cephalosporin C biosynthesis. Downregulation of pcbC can be detected easily by a bioassay measuring the antibiotic activity of individual strains. In addition, the presence of the isopenicillin N synthase was investigated by Western blot hybridization. All transformants having a colorless phenotype showed simultaneous downregulation of the pcbC gene, albeit at different levels. The RNA-silencing system presented here should be a powerful genetic tool for strain improvement and genome-wide analysis of this biotechnologically important filamentous fungus.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.