A dark-to-bright reporter cell for classical swine fever virus infection

Antiviral Research
Fuwang ChenLinzhu Ren

Abstract

Current methods to quantitate classical swine fever virus (CSFV) infectivity in cell culture are time-consuming and labor-intensive. This study described the generation of a dark-to-bright fluorescent reporter cells to facilitate in vitro studies of CSFV infection and replication. This assay was based on a novel reporter cell stably expressing the enhanced green fluorescent protein (EGFP) fused in-frame to a quenching peptide via a special recognition sequence of the CSFV NS3 protease. Chromophore maturation of EGFP can be prevented by quenching peptide until the quenching peptide was specifically cleaved by NS3 protease during CSFV infection, making it a dark-to-bright reporter of CSFV infection. The result demonstrated that the CSFV-infected cells were clearly distinguishable from mock-infected cells and cells infected with other viruses. There was a strong correlation between the fluorescence intensity and viral RNA replication in CSFV-infected cells. The cell enabled rapid and sensitive detection of CSFV infection and viral replication in cell culture. The best time to examine the fluorescence in CSFV-infected cells was at 48h post-inoculation. These data suggested that the cells can be used as a reporter cell in CSFV infec...Continue Reading

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Citations

Jan 6, 2016·Applied Biochemistry and Biotechnology·Linzhu RenHongsheng Ouyang
Aug 11, 2016·Virus Research·Zhiyuan PengLinzhu Ren
Jul 3, 2021·Viruses·Carina Maria ReuscherBenjamin Lamp

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Antivirals are medications that are used specifically for treating viral infections. Discover the latest research on antivirals here.