Abstract：
:Human immunodeficiency virus (HIV) encodes a transcription trans-activator (Tat) with an essential role in the transcriptional elongation of viral RNA based on the viral promoter long terminal repeat (LTR). Tat-mediated transcription is conserved and can be distinguished from host transcription, so it is a therapeutic target for combating HIV replication. Traditional screening assays for Tat-mediated transcriptional inhibitors are based on the biochemical properties of Tat and transactivation-responsive RNA. We developed an inducible system based on two lentiviral expression cassettes for doxycycline (Dox)-inducible Tat and Renilla luciferase (R-Luc) using TZM-bl cells harboring LTR-driven firefly luciferase (F-Luc). The cells simultaneously expressed both Tat-induced F-Luc and R-Luc, so it was possible to recognize off-target effects in the presence of Dox. The system was validated with known inhibitors: CYC202 obtained high sensitivity and specificity, whereas 6Bio and DRB had off-target effects. The MTT-based cytotoxicity test indicated the resistance of the system even at concentrations with off-target effects. The specificity of the system was confirmed using antiretroviral drugs. Our dual reporter system can simply detect Tat inhibitory effects, as well as precisely discriminate between the inhibitory and off-target effects of inhibitors, and may be useful for the development of a therapeutic anti-HIV drug.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Shin Y,Choi BS,Kim KC,Kang C,Kim K,Yoon CHdoi
10.1016/j.jviromet.2017.08.005subject
Has Abstractpub_date
2017-11-01 00:00:00pages
1-9eissn
0166-0934issn
1879-0984pii
S0166-0934(17)30143-Xjournal_volume
249pub_type
杂志文章abstract：:A solid phase reverse passive hemadsorption test (SP-RPHAd) for hepatitis B surface antigen detection is described. It was compared with a commercial reverse passive hemagglutination assay (Hepatest, Wellcome, U.K.). SP-RPHAd is four-fold less expensive than Hepatest and undiluted sera can be used instead of eight-fol...
journal_title：Journal of virological methods
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journal_title：Journal of virological methods
pub_type： 杂志文章
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pub_type： 杂志文章
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更新日期：2008-09-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
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更新日期：2008-11-01 00:00:00
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更新日期：2012-01-01 00:00:00
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pub_type： 杂志文章
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journal_title：Journal of virological methods
pub_type： 临床试验,杂志文章
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更新日期：2006-09-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
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journal_title：Journal of virological methods
pub_type： 杂志文章
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更新日期：2010-11-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
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journal_title：Journal of virological methods
pub_type： 杂志文章
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更新日期：2001-04-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(90)90107-q
更新日期：1990-08-01 00:00:00
