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Journal of Virology, July 2005, p. 8942-8947, Vol. 79, No. 14
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.14.8942-8947.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Nairovirus RNA Sequences Expressed by a Semliki Forest Virus Replicon Induce RNA Interference in Tick Cells

Stephan Garcia,1,2 Agnès Billecocq,1 Jean-Marc Crance,2 Ulrike Munderloh,3 Daniel Garin,2 and Michèle Bouloy1*

Unité de Génétique moléculaire des Bunyaviridés, Institut Pasteur, Paris, France,1 Laboratoire de virologie, CRSSA, Centre de Recherche Emile Pardé, Grenoble, France,2 University of Minnesota, Department of Entomology, St. Paul, Minnesota 551083

Received 4 February 2005/ Accepted 7 April 2005

We report the successful infection of the cell line ISE6 derived from Ixodes scapularis tick embryos by the tick-borne Hazara virus (HAZV), a nairovirus in the family Bunyaviridae. Using a recombinant Semliki Forest alphavirus replicon that replicates in these cells, we were able to inhibit replication of HAZV, and we showed that this blockage is mediated by the replication of the Semliki Forest alphavirus replicon; the vector containing the HAZV nucleoprotein gene in sense or antisense orientation efficiently inhibited HAZV replication. Moreover, expression of a distantly related nucleoprotein gene from Crimean-Congo hemorrhagic fever nairovirus failed to induce HAZV silencing, indicating that the inhibition is sequence specific. The resistance of these cells to replicate HAZV correlated with the detection of specific RNase activity and 21- to 24-nucleotide-long small interfering RNAs. Altogether, these results strongly suggest that pathogen-derived resistance can be established in the tick cells via a mechanism of RNA interference.

* Corresponding author. Mailing address: Institut Pasteur, Unité de Génétique Moléculaire des Bunyaviridés, 25 rue du Dr Roux, Paris 75015, France. Phone: 33 1 40 61 31 57. Fax: 33 1 40 61 31 51. E-mail: mbouloy{at}pasteur.fr.

Journal of Virology, July 2005, p. 8942-8947, Vol. 79, No. 14
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.14.8942-8947.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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