lunes, 21 de abril de 2014

High-throughput, luciferase-based reverse gene... [Antiviral Res. 2014] - PubMed - NCBI

High-throughput, luciferase-based reverse gene... [Antiviral Res. 2014] - PubMed - NCBI

 2014 Apr 5. pii: S0166-3542(14)00092-8. doi: 10.1016/j.antiviral.2014.03.018. [Epub ahead of print]

High-throughput, luciferase-based reverse genetics systems for identifying inhibitors of Marburg and Ebola viruses.


Marburg virus (MARV) and Ebola virus (EBOV), members of the family Filoviridae, represent a significant challenge to global public health. Currently, no licensed therapies exist to treat filovirus infections, which cause up to 90% mortality in human cases. To facilitate development of antivirals against these viruses, we established two distinct screening platforms based on MARV and EBOV reverse genetics systems that express secreted Gaussia luciferase (gLuc). The first platform is a mini-genome replicon to screen viral replication inhibitors using gLuc quantification in a BSL-2 setting. The second platform is complementary to the first and expresses gLuc as a reporter gene product encoded in recombinant infectious MARV and EBOV, thereby allowing for rapid quantification of viral growth during treatment with antiviral compounds. We characterized these viruses by comparing luciferase activity to virus production, and validated luciferase activity as an authentic real-time measure of viral growth. As proof of concept, we adapt both mini-genome and infectious virus platforms to high-throughput formats, and demonstrate efficacy of several antiviral compounds. We anticipate that both approaches will prove highly useful in the development of anti-filovirus therapies, as well as in basic research on the filovirus life cycle.
Published by Elsevier B.V.


Antiviral screen, Ebola virus, Filovirus, Luciferase, Marburg virus, Reverse genetics

[PubMed - as supplied by publisher]

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