Reference : RNA silencing in the dermatophyte Microsporum canis
Scientific journals : Article
Life sciences : Microbiology
http://hdl.handle.net/2268/9063
RNA silencing in the dermatophyte Microsporum canis
English
Vermout, S. [> > > >]
Tabart, J. [> > > >]
Baldo, Aline mailto [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Parasitologie et pathologie des maladies parasitaires >]
Monod, M. [> > > >]
Losson, Bertrand mailto [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Parasitologie et pathologie des maladies parasitaires >]
Mignon, Bernard mailto [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Parasitologie et pathologie des maladies parasitaires >]
Oct-2007
FEMS Microbiology Letters
Blackwell Publishing
275
1
38-45
Yes (verified by ORBi)
International
0378-1097
Oxford
[en] Microsporum canis ; dermatophyte ; RNA silencing ; SUB3 ; DPPIV
[en] Dermatomycoses caused by Microsporum canis are frequent in domestic animals and easily transmissible to humans. Several proteases secreted by this fungus were identified as potential virulence factors, but the construction of deficient strains is required to investigate their role in the pathogenesis of the disease. Using target genes encoding two of these proteases, a first evaluation of the utility of RNA-mediated silencing as a reverse genetic tool in dermatophytes was carried out. SUB3 and DPPIV, respectively coding for a subtilisin and a dipeptidyl peptidase, were both down-regulated, by means of two plasmid constructs designed to express an RNA hairpin that corresponds to part of their respective sequence. The degree of attenuation was evaluated by enzymatic assay of the transformants culture supernatants, and by real-time reverse transcriptase-polymerase chain reaction. Enzymatic activities and expression levels varied from less than 5% to 100% of that of control transformants obtained with plasmid without hairpin inserts. Inhibition was globally more efficient for SUB3 than for DPPIV. These results show that RNA silencing can be used for functional genomics in M. canis, and particularly to circumvent the limits and technical difficulties of conventional disruption methods.
http://hdl.handle.net/2268/9063
10.1111/j.1574-6968.2007.00870.x

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