Recovery of an arenavirus entirely from RNA polymerase I/II-driven cDNA

Proceedings of the National Academy of Sciences of the United States of America
Lukas FlatzDaniel D Pinschewer

Abstract

The prototypic arenavirus lymphocytic choriomeningitis virus has been a primary workhorse of viral immunologists for almost a century, and it has served as an important model for studying basic principles of arenavirus molecular biology. Its negative-stranded bisegmented RNA genome has, however, posed a major obstacle to attempts at manipulating the infectious virus by reverse genetic techniques. Here, we report the recovery of infectious lymphocytic choriomeningitis virus (the immunosuppressive strain clone 13) entirely from cDNA. Intracellular transcription of the short and the long viral genome segment from polymerase (pol) I-driven vectors and coexpression of the minimal viral-transacting factors NP and L from pol II-driven plasmids resulted in the efficient formation of infectious virus with genetic tags in both genome segments. The cDNA-derived viruses behaved identically to wild-type virus in both cell culture and infected mice. Importantly, they caused a chronic infection and suppressed the adaptive immune response to an unrelated third-party virus. This technology provides an important basis for investigating viral determinants of persistent infection and immunosuppression. In addition, our findings demonstrate that po...Continue Reading

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Citations

Feb 12, 2009·Proceedings of the National Academy of Sciences of the United States of America·Sebastien F EmonetJuan C de la Torre
Nov 26, 2010·Proceedings of the National Academy of Sciences of the United States of America·Andreas BergthalerDaniel D Pinschewer
Jun 30, 2010·International Immunology·Daniel D PinschewerAndreas Bergthaler
Feb 11, 2011·Nucleic Acids Research·Takeshi YoshidaKiyohito Yagi
Dec 1, 2010·Antimicrobial Agents and Chemotherapy·Michelle MendenhallBrian B Gowen
Apr 12, 2013·Journal of Virology·Patrick HeinemannStephan Günther
Sep 7, 2007·Journal of Virology·Luis Martínez-SobridoJuan Carlos de la Torre
Apr 10, 2009·PLoS Biology·Andreas BergthalerDaniel D Pinschewer
Jan 12, 2013·PloS One·Cyrille J BonhommeMichael J Buchmeier
Jun 6, 2006·PLoS Pathogens·Andreas BergthalerDaniel D Pinschewer
Jan 25, 2013·Viruses·Juan C Zapata, Maria S Salvato
Aug 2, 2013·Journal of Virology·Nadezhda E YunSlobodan Paessler
Jun 27, 2014·Viruses·Christopher C Stobart, Martin L Moore
May 23, 2012·Expert Opinion on Drug Discovery·Sheli R RadoshitzkySina Bavari
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Jul 23, 2016·Viruses·Luis Martínez-Sobrido, Juan Carlos de la Torre
Jul 28, 2016·The Journal of Experimental Medicine·Daniel T UtzschneiderDietmar Zehn

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