RNA silencing in the dermatophyte Microsporum canis

[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.

Disciplines :

Microbiology

Author, co-author :

Vermout, S.

Tabart, J.

Baldo, Aline ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Parasitologie et pathologie des maladies parasitaires

Monod, M.

Losson, Bertrand ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Parasitologie et pathologie des maladies parasitaires

Mignon, Bernard ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Parasitologie et pathologie des maladies parasitaires

Language :

English

Title :

RNA silencing in the dermatophyte Microsporum canis

Publication date :

October 2007

Journal title :

FEMS Microbiology Letters

ISSN :

0378-1097

eISSN :

1574-6968

Publisher :

Blackwell Publishing, Oxford, United Kingdom

Volume :

275

Issue :

Pages :

38-45

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 17 March 2009

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