Efficient gene silencing by expression of double stranded RNA in Neurospora crassa

Goldoni, Marina; Azzalin, Gianluca; Macino, Giuseppe; Cogoni, Carlo

doi:10.1016/j.fgb.2004.08.002

Cited by 108 publications

(98 citation statements)

References 24 publications

(41 reference statements)

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“…Longer stem hairpins (ca. 500 bp) have been used in several other fungi (Rappleye et al, 2004;Goldoni et al, 2004;Tanguay et al, 2006), and we had previously observed that shorter hairpins (162 bp) could initiate down-regulation of GFP in C. cinerea (Heneghan et al, 2007).…”

Section: Resultsmentioning

confidence: 99%

Oligonucleotide sequences forming short self-complimentary hairpins can expedite the down-regulation of Coprinopsis cinerea genes

Costa

Mills

Bailey

et al. 2008

Journal of Microbiological Methods

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This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. This technological advance should expedite functional genomic studies in C. cinerea and has wider potential for utility in other homobasidiomycete and filamentous fungi. A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT

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Section: Resultsmentioning

confidence: 99%

Oligonucleotide sequences forming short self-complimentary hairpins can expedite the down-regulation of Coprinopsis cinerea genes

Costa

Mills

Bailey

et al. 2008

Journal of Microbiological Methods

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“…In its most effective form, an ectopically integrated RNAi construct codes for double stranded RNA, identical to part of the endogenous sequence being targeted [120,[123][124][125]. The linear RNAi knockdown transcript contains an inverse repeat of the target sequence that folds up into a "hairpin" in which the inverse repeat is double stranded.…”

Section: Post-transcriptional Gene Silencingmentioning

confidence: 99%

“…The repeat sequences are typically about 500 bp of coding sequence separated by a spacer sequence of about 250 bp [126]. A substantial decrease in efficiency was found when using repeat sequences of 200 bp rather than 600 bp or 900 bp [125]. The inclusion of an intron in the spacer region may greatly increase the silencing efficiency, possibly through enhanced export of the hairpin from the nucleus during splicing [125].…”

Section: Post-transcriptional Gene Silencingmentioning

confidence: 99%

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Approaches to functional genomics in filamentous fungi

et al. 2006

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The study of gene function in filamentous fungi is a field of research that has made great advances in very recent years. A number of transformation and gene manipulation strategies have been developed and applied to a diverse and rapidly expanding list of economically important filamentous fungi and oomycetes. With the significant number of fungal genomes now sequenced or being sequenced, functional genomics promises to uncover a great deal of new information in coming years. This review discusses recent advances that have been made in examining gene function in filamentous fungi and describes the advantages and limitations of the different approaches.

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“…Since then, many vectors for RNA-mediated gene silencing in fungi have been used successfully (Goldoni et al 2004;Fitzgerald et al 2004;Mouyna et al 2004;Nakayashiki et al 2005). Interestingly, there is indication of occurrence of small interfering RNA-mediated mechanisms in A. niger (Barnes et al 2008).…”

Section: Introductionmentioning

confidence: 99%

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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Efficient gene silencing by expression of double stranded RNA in Neurospora crassa

Cited by 108 publications

References 24 publications

Oligonucleotide sequences forming short self-complimentary hairpins can expedite the down-regulation of Coprinopsis cinerea genes

Oligonucleotide sequences forming short self-complimentary hairpins can expedite the down-regulation of Coprinopsis cinerea genes

Approaches to functional genomics in filamentous fungi

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

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