RNA Silencing in Filamentous Fungi: From Basics to Applications

RNA silencing approaches for identifying pathogenicity and virulence elements towards engineering crop resistance to plant pathogenic fungi.

“…This ability can be specifically useful to fungi with low gene targeting efficiency or those which are multi-nucleated. Simultaneous silencing of homologous genes has been achieved by targeting a conserved sequence of a gene family [32]; (2) RNAi does not change the genomic structure of target genes and is extremely useful in studies when deletion of target genes is lethal for the recipient since it does not completely shut down gene expression; (3) Since the efficiency of homologous recombination varies considerably among fungal species, RNAi can offer a convenient genomic tool in systems where the efficiency of gene targeting is low; (4) RNAi requires only short stretches of sequence information and thus is a major advantage for organisms for which there is little genome information available; (5) The efficiency of RNAi is not compromised by the presence of non-transformed nuclei or multi-copy genes due to aneuploidy [3]. Most fungi possess multinucleated or multicellular hyphae and some exhibit heterokaryosis making gene targeting inefficient and complicated; (6) By making use of inducible promoters, RNA silencing can facilitate the study of gene expression at specific developmental stages or in different parts of the organism.…”

“…One powerful technology to assess gene function exploits posttranscriptional gene silencing (PTGS) mechanisms, which act through various RNA intermediates [1,2]. This RNAmediated gene silencing, referred to as RNA interference or RNAi, has been shown many times over to be an excellent tool for gene targeting to study their role in plant disease caused by fungal [3,4], bacterial [5] and viral [6] pathogens. The molecular toolbox for modulating gene functions in PPF and oomycetes has considerably enlarged over the last few years.…”

Peer Review #1 of "Development of an RNA interference (RNAi) gene knockdown protocol in the anaerobic gut fungus Pecoramyces ruminantium strain C1A (v0.1)"

“…The role played by non-coding RNA (ncRNA) molecules in epigenetic modulation of gene expression at the transcriptional and post-transcriptional levels is now well recognized (1). Small interfering RNAs (siRNA) are short (20-24 nt) double stranded RNA molecules that mediate post-transcriptional regulation of gene expression and gene silencing by binding to mRNA in a sequence-specific manner (2). The process of RNA interference (RNAi) has been independently documented in fungi (3)(4)(5), animals and human cell lines (6,7), as well as plants (8).…”

“…Manuscript to be reviewed 19). On the other hand, the introduction of foreign siRNA could be utilized for targeted, sequence-specific, gene bnocbdown in fungi (2,3,5). Indeed, demonstration of the feasibility of RNAi approaches for targeted gene silencing has been shown in Ascomycota (5,(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30),…”

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“…Under normal physiological conditions, RNAi is thought to play a role in endogenous regulation of gene expression (13), development of resistance to viruses (14)(15)(16)(17), and silencing the expression of transposons (18,19). On the other hand, the introduction of foreign siRNA could be utilized for targeted, sequence-specific, gene knockdown in fungi (2,3,5). Indeed, demonstration of the feasibility of RNAi approaches for targeted gene silencing has been shown in Ascomycota (5,(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30),…”