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Proc Natl Acad Sci USA. Dengue reporter viruses reveal viral dynamics in interferon receptor-deficient mice and sensitivity to interferon effectors in vitro
[Source: Proceedings of the National Academy of the Science of the United States of America, full page: (LINK). Abstract, edited.]
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Dengue reporter viruses reveal viral dynamics in interferon receptor-deficient mice and sensitivity to interferon effectors in vitro
John W. Schoggins, Marcus Dorner, Michael Feulner, Naoko Imanaka, Mary Y. Murphy, Alexander Ploss, and Charles M. Rice<SUP>1</SUP>
<SUP></SUP>
Author Affiliations: Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10065
Contributed by Charles M. Rice, July 19, 2012 (sent for review June 16, 2012)
Abstract
Dengue virus (DENV) is a global disease threat for which there are no approved antivirals or vaccines. Establishing state-of-the-art screening systems that rely on fluorescent or luminescent reporters may accelerate the development of anti-DENV therapeutics. However, relatively few reporter DENV platforms exist. Here, we show that DENV can be genetically engineered to express a green fluorescent protein or firefly luciferase. Reporter viruses are infectious in vitro and in vivo and are sensitive to antiviral compounds, neutralizing antibodies, and interferons. Bioluminescence imaging was used to follow the dynamics of DENV infection in mice and revealed that the virus localized predominantly to lymphoid and gut-associated tissues. The high-throughput potential of reporter DENV was demonstrated by screening a library of more than 350 IFN-stimulated genes for antiviral activity. Several antiviral effectors were identified, and they targeted DENV at two distinct life cycle steps. These viruses provide a powerful platform for applications ranging from validation of vaccine candidates to antiviral discovery.
in vivo imaging / antiviral screening / flavivirus / interferon-stimulated gene
Footnotes
<SUP>1</SUP>To whom correspondence should be addressed. E-mail: ricec@rockefeller.edu.
Author contributions: J.W.S., M.D., and C.M.R. designed research; J.W.S., M.D., M.F., N.I., and M.Y.M. performed research; M.D. and A.P. contributed new reagents/analytic tools; J.W.S., M.D., and C.M.R. analyzed data; and J.W.S., M.D., A.P., and C.M.R. wrote the paper.
The authors declare no conflict of interest.
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1212379109/-/DCSupplemental.
-John W. Schoggins, Marcus Dorner, Michael Feulner, Naoko Imanaka, Mary Y. Murphy, Alexander Ploss, and Charles M. Rice<SUP>1</SUP>
<SUP></SUP>
Author Affiliations: Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10065
Contributed by Charles M. Rice, July 19, 2012 (sent for review June 16, 2012)
Abstract
Dengue virus (DENV) is a global disease threat for which there are no approved antivirals or vaccines. Establishing state-of-the-art screening systems that rely on fluorescent or luminescent reporters may accelerate the development of anti-DENV therapeutics. However, relatively few reporter DENV platforms exist. Here, we show that DENV can be genetically engineered to express a green fluorescent protein or firefly luciferase. Reporter viruses are infectious in vitro and in vivo and are sensitive to antiviral compounds, neutralizing antibodies, and interferons. Bioluminescence imaging was used to follow the dynamics of DENV infection in mice and revealed that the virus localized predominantly to lymphoid and gut-associated tissues. The high-throughput potential of reporter DENV was demonstrated by screening a library of more than 350 IFN-stimulated genes for antiviral activity. Several antiviral effectors were identified, and they targeted DENV at two distinct life cycle steps. These viruses provide a powerful platform for applications ranging from validation of vaccine candidates to antiviral discovery.
in vivo imaging / antiviral screening / flavivirus / interferon-stimulated gene
Footnotes
<SUP>1</SUP>To whom correspondence should be addressed. E-mail: ricec@rockefeller.edu.
Author contributions: J.W.S., M.D., and C.M.R. designed research; J.W.S., M.D., M.F., N.I., and M.Y.M. performed research; M.D. and A.P. contributed new reagents/analytic tools; J.W.S., M.D., and C.M.R. analyzed data; and J.W.S., M.D., A.P., and C.M.R. wrote the paper.
The authors declare no conflict of interest.
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1212379109/-/DCSupplemental.
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