Quelling: transient inactivation of gene expression in <i>Neurospora crassa</i> by transformation with homologous sequences

Romano, Nicoletta; Macino, Giuseppe

doi:10.1111/j.1365-2958.1992.tb02202.x

Cited by 700 publications

(467 citation statements)

References 32 publications

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“…Such approach also holds huge therapeutic potential in modern pharmacologic applications [4,[6][7][8][9][10]. Antisense oligonucleotides either induce RNase H activity or cause steric blocking that physically disturbs splicing or even the whole translational machinery [11,12].…”

Section: Introductionmentioning

confidence: 99%

Tandem Mass Spectrometry of Modified and Platinated Oligoribonucleotides

Nyakas

Stucki

Schürch

2011

J. Am. Soc. Mass Spectrom.

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Therapeutic approaches for treatment of various diseases aim at the interruption of transcription or translation. Modified oligonucleotides, such as 2′-O-methyl-and methylphosphonatederivatives, exhibit high resistance against cellular nucleases, thus rendering application for, e.g., antigene or antisense purposes possible. Other approaches are based on administration of cross-linking agents, such as cis-diamminedichloroplatinum(II) (cisplatin, DDP), which is still the most widely used anticancer drug worldwide. Due to the formation of 1,2-intrastrand cross links at adjacent guanines, replication of the double-strand is disturbed, thus resulting in significant cytotoxicity. Evidence for the gas-phase dissociation mechanism of platinated RNA is given, based on nano-electrospray ionization high-resolution multistage tandem mass spectrometry (MS n ). Confirmation was found by investigating the fragmentation pattern of platinated and unplatinated 2′-methoxy oligoribonucleotide hexamers and their corresponding methylphosphonate derivatives. Platinated 2′-methoxy oligoribonucleotides exhibit a similar gas-phase dissociation behavior as the corresponding DNA and RNA sequences, with the 3′-C-O bond adjacent to the vicinal guanines being cleaved preferentially, leading to w x -ion formation. By examination of the corresponding platinated methylphosphonate derivatives of the 2′-methoxy oligoribonucleotides, the key role of the negatively charged phosphate oxygen atoms in direct proximity to the guanines was proven. The significant alteration of fragmentation due to platination is demonstrated by comparison of the fragment ion patterns of unplatinated and platinated 2′-O-methyl-and 2′-O-methyl methylphosphonate oligoribonucleotides, and the results obtained by H/D exchange experiments.

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Section: Introductionmentioning

confidence: 99%

Tandem Mass Spectrometry of Modified and Platinated Oligoribonucleotides

Nyakas

Stucki

Schürch

2011

J. Am. Soc. Mass Spectrom.

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“…Since the discovery of the RNAi phenomenon in Neurospora crassa (Romano & Macino, 1992), several studies have provided evidence for RNA silencing machineries in various filamentous fungi and yeasts (reviewed by . Comprehensive phylogenetic analyses of diverse fungal genomes revealed a difference in the number of paralogous RNAsilencing proteins, such as Argonautes, Dicers and RNAdependent RNA polymerases (Nakayashiki et al, 2006;.…”

Section: Introductionmentioning

confidence: 99%

Evidence for Dicer-dependent RNA interference in the industrial penicillin producer Penicillium chrysogenum

2009

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RNA interference (RNAi) is a sequence-specific post-transcriptional gene silencing system that downregulates target gene expression. Here, we provide several lines of evidence for RNA silencing in the industrial β-lactam antibiotic producer Penicillium chrysogenum using the DsRed reporter gene under the control of the constitutive trpC promoter or the inducible xylP promoter. The functional RNAi system was verified by detection of siRNAs that hybridized exclusively with gene-specific 32P-labelled RNA probes. Moreover, when RNAi was used to silence the endogenous PcbrlA morphogene that controls conidiophore development, a dramatic reduction in the formation of conidiospores was observed in 47 % of the corresponding transformants. Evidence that RNAi in P. chrysogenum is dependent on a Dicer peptide was provided with a strain lacking Pcdcl2. In the ΔPcdcl2 background, silencing of the PcbrlA gene was tested. None of the transformants analysed showed a developmental defect. The applicability of the RNAi system in P. chrysogenum was finally demonstrated by silencing the Pcku70 gene to increase homologous recombination frequency. This led to the generation of single and double knockout mutants.

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“…Fungal RNAi mechanisms were first identified in the saprotrophic species Neurospora crassa and termed ‘quelling’ 19. Quelling is active in the vegetative phase of the N. crassa life cycle and is necessary to control transposons 20.…”

Section: Rna Mechanismsmentioning

confidence: 99%

RNAi as an emerging approach to control Fusarium head blight disease and mycotoxin contamination in cereals

Machado

Brown

Urban

et al. 2017

Pest Management Science

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Fusarium graminearum is a major fungal pathogen of cereals worldwide, causing seedling, stem base and floral diseases, including Fusarium head blight (FHB). In addition to yield and quality losses, FHB contaminates cereal grain with mycotoxins, including deoxynivalenol, which are harmful to human, animal and ecosystem health. Currently, FHB control is only partially effective due to several intractable problems. RNA interference (RNAi) is a natural mechanism that regulates gene expression. RNAi has been exploited in the development of new genomic tools that allow the targeted silencing of genes of interest in many eukaryotes. Host‐induced gene silencing (HIGS) is a transgenic technology used to silence fungal genes in planta during attempted infection and thereby reduces disease levels. HIGS relies on the host plant's ability to produce mobile small interfering RNA molecules, generated from long double‐stranded RNA, which are complementary to targeted fungal genes. These molecules are transferred from the plant to invading fungi via an uncharacterised mechanism, to cause gene silencing. Here, we describe recent advances in RNAi‐mediated control of plant pathogenic fungi, highlighting the key advantages and disadvantages. We then discuss the developments and implications of combining HIGS with other methods of disease control. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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Quelling: transient inactivation of gene expression in Neurospora crassa by transformation with homologous sequences

Cited by 700 publications

References 32 publications

Tandem Mass Spectrometry of Modified and Platinated Oligoribonucleotides

Tandem Mass Spectrometry of Modified and Platinated Oligoribonucleotides

Evidence for Dicer-dependent RNA interference in the industrial penicillin producer Penicillium chrysogenum

RNAi as an emerging approach to control Fusarium head blight disease and mycotoxin contamination in cereals

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