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Journal of Virology, June 2009, p. 5606-5614, Vol. 83, No. 11
0022-538X/09/$08.00+0     doi:10.1128/JVI.00276-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Efficient Reverse Genetics Generation of Infectious Junin Viruses Differing in Glycoprotein Processing{triangledown} ,{dagger}

César G. Albariño,1 Éric Bergeron,1 Bobbie Rae Erickson,1 Marina L. Khristova,2 Pierre E. Rollin,1 and Stuart T. Nichol1*

Special Pathogens Branch,1 Biotechnology Core Facility Branch, Centers for Disease Control and Prevention, 1600 Clifton Road, MS G-14, Atlanta, Georgia 303332

Received 7 February 2009/ Accepted 11 March 2009

The New World arenaviruses, Junin, Machupo, Guanarito, Sabia, and Chapare, are associated with rapidly progressing severe hemorrhagic fever with a high rate of case fatality in various regions of South America. The threat of natural or deliberate outbreaks associated with these viruses makes the development of preventive or therapeutic measures important. Here we describe a Junin virus functional minigenome system and a reverse genetics system for production of infectious Junin virus. This robust, highly efficient system involves transfection of cells with only two plasmids which transcribe the virus S and L antigenomic RNAs. The utility of the system is demonstrated by generating Junin viruses which encode a glycoprotein precursor (GPC) containing the following: (i) the wild-type (SKI-1/S1P peptidase) cleavage site, (ii) no cleavage site, or (iii) a cleavage site where the SKI-1/S1P motif (RSLK) is replaced by a furin cleavage site (RRKR). In contrast to the wild-type virus, Junin virus lacking a GPC cleavage site replicated within successfully transfected cells but failed to yield infectious virus particles. This confirms observations with other arenaviruses suggesting that GPC cleavage is essential for arenavirus infectivity. In contrast, infectious Junin virus which encoded GPC cleaved by furin-like proteases was easily generated. The two-plasmid, high efficiency aspects of this Junin virus reverse genetics system show great promise for addressing important questions regarding arenavirus hemorrhagic fever disease and for development of precisely attenuated live arenavirus vaccines.

* Corresponding author. Mailing address: Special Pathogens Branch, Centers for Disease Control and Prevention, 1600 Clifton Road, MS G-14, Atlanta, GA 30333. Phone: (404) 639-1115. Fax: (404) 639-1118. E-mail: stn1{at}cdc.gov

{triangledown} Published ahead of print on 25 March 2009.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, June 2009, p. 5606-5614, Vol. 83, No. 11
0022-538X/09/$08.00+0     doi:10.1128/JVI.00276-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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