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Information Journal Paper

Title

TOWARD THE DEVELOPMENT OF A SINGLE-ROUND INFECTION ASSAY BASED ON EGFP REPORTING FOR ANTI-HIV-1 DRUG DISCOVERY

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Abstract

 Background: The rapid increase of HIV-1 strains resistant to current antiretroviral drugs is a challenge for successful AIDS therapy. This necessitates the development of novel drugs, and to this end, availability of screening systems for in vitro DRUG DISCOVERY is a priority. Herein, we report the modification of a previously developed system for increased sensitivity, ease of use, and costefficiency, based on the application of the EGFP marker.Methods: A PCR-amplified gfp gene (gfp) was cloned into pmzNL4-3, the plasmid already designed to produce single-cycle replicable virions, in frame with the reverse-transcriptase gene to construct the pmzNL4-3/GFP plasmid. GFP-mzNL4-3 pseudo-typed virions, as the first progeny viruses, were recovered from the culture supernatant of HEK293T cells co-transfected with pmzNL4-3/GFP and the helper plasmids pSPAX2 and pMD2G, which respectively encode HIV-1 Gag-Pol and vesicular stomatitis virus glycoprotein. Single-cycle replication and virion production were assessed by syncytia formation, p24 antigen assays, and electron and fluorescence microscopy.Results: The incorporation of EGFP into the viral particles allowed their quantification by FLUOROMETRY, flow-cytometry, and fluorescence microscopy; however, this modification did not affect the single-round infectivity or production rate of the GFP fluorescence-emitting virions.Conclusions: Our results certify the development of a rapid, inexpensive, and safe GFP-reporting single-cycle replicable system for anti-HIV DRUG DISCOVERY. Further experiments are needed to measure the validity and robustness of the assay.

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    APA: Copy

    SOEZI, MAHDIEH, MEMARNEJADIAN, ARASH, AMINZADEH, SAEED, ZABIHOLLAHI, REZVAN, SADAT, SEYED MEHDI, AMINI, SAFIEH, HEKMAT, SOHEILA, & AGHASADEGHI, MOHAMMAD REZA. (2015). TOWARD THE DEVELOPMENT OF A SINGLE-ROUND INFECTION ASSAY BASED ON EGFP REPORTING FOR ANTI-HIV-1 DRUG DISCOVERY. REPORTS OF BIOCHEMISTRY AND MOLECULAR BIOLOGY, 4(1), 0-0. SID. https://sid.ir/paper/343092/en

    Vancouver: Copy

    SOEZI MAHDIEH, MEMARNEJADIAN ARASH, AMINZADEH SAEED, ZABIHOLLAHI REZVAN, SADAT SEYED MEHDI, AMINI SAFIEH, HEKMAT SOHEILA, AGHASADEGHI MOHAMMAD REZA. TOWARD THE DEVELOPMENT OF A SINGLE-ROUND INFECTION ASSAY BASED ON EGFP REPORTING FOR ANTI-HIV-1 DRUG DISCOVERY. REPORTS OF BIOCHEMISTRY AND MOLECULAR BIOLOGY[Internet]. 2015;4(1):0-0. Available from: https://sid.ir/paper/343092/en

    IEEE: Copy

    MAHDIEH SOEZI, ARASH MEMARNEJADIAN, SAEED AMINZADEH, REZVAN ZABIHOLLAHI, SEYED MEHDI SADAT, SAFIEH AMINI, SOHEILA HEKMAT, and MOHAMMAD REZA AGHASADEGHI, “TOWARD THE DEVELOPMENT OF A SINGLE-ROUND INFECTION ASSAY BASED ON EGFP REPORTING FOR ANTI-HIV-1 DRUG DISCOVERY,” REPORTS OF BIOCHEMISTRY AND MOLECULAR BIOLOGY, vol. 4, no. 1, pp. 0–0, 2015, [Online]. Available: https://sid.ir/paper/343092/en

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