Journal
VIROLOGY
Volume 486, Issue -, Pages 78-87Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.virol.2015.08.029
Keywords
Retrovirus; HIV; Assembly; Assay; Inhibitor; High-throughput screening; Capsid; CA
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Funding
- Grant Agency of the Czech Republic [14-15326S]
- NPU I Projects from Ministry of Education [LO 1302, LO 1304, LN12011]
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Due to the high number of drug-resistant HIV-1 mutants generated by highly active antiretroviral therapy (HAART), there is continuing demand for new types of inhibitors. Both the assembly of the Gag polyprotein into immature and mature HIV-1 particles are attractive candidates for the blocking of the retroviral life cycle. Currently, no therapeutically-used assembly inhibitor is available. One possible explanation is the lack of a reliable and simple assembly inhibitor screening method. To identify compounds potentially inhibiting the formation of both types of HIV-1 particles, we developed a new fluorescent high-throughput screening assay. This assay is based on the quantification of the assembly efficiency in vitro in a 96-well plate format. The key components of the assay are HIV-1 Gag-derived proteins and a dual-labelled oligonucleotide, which emits fluorescence only when the assembly of retroviral particles is inhibited. The method was validated using three (CAI, BM2, FF74) reported assembly inhibitors. (C) 2015 The Authors. Published by Elsevier Inc.
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