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A new system for parallel drug screening against multiple-resistant HIV mutants based on lentiviral self-inactivating (SIN) vectors and multi-colour analyses

BACKGROUND: Despite progress in the development of combined antiretroviral therapies (cART), HIV infection remains a significant challenge for human health. Current problems of cART include multi-drug-resistant virus variants, long-term toxicity and enormous treatment costs. Therefore, the identific...

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Autores principales: Prokofjeva, Maria M, Riecken, Kristoffer, Spirin, Pavel V, Yanvarév, Dimitriy V, Düsedau, Arne, Ellinger, Bernhard, Fehse, Boris, Stocking, Carol, Prassolov, Vladimir S
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3560153/
https://www.ncbi.nlm.nih.gov/pubmed/23286882
http://dx.doi.org/10.1186/1742-6405-10-1
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author Prokofjeva, Maria M
Riecken, Kristoffer
Spirin, Pavel V
Yanvarév, Dimitriy V
Düsedau, Arne
Ellinger, Bernhard
Fehse, Boris
Stocking, Carol
Prassolov, Vladimir S
author_facet Prokofjeva, Maria M
Riecken, Kristoffer
Spirin, Pavel V
Yanvarév, Dimitriy V
Düsedau, Arne
Ellinger, Bernhard
Fehse, Boris
Stocking, Carol
Prassolov, Vladimir S
author_sort Prokofjeva, Maria M
collection PubMed
description BACKGROUND: Despite progress in the development of combined antiretroviral therapies (cART), HIV infection remains a significant challenge for human health. Current problems of cART include multi-drug-resistant virus variants, long-term toxicity and enormous treatment costs. Therefore, the identification of novel effective drugs is urgently needed. METHODS: We developed a straightforward screening approach for simultaneously evaluating the sensitivity of multiple HIV gag-pol mutants to antiviral drugs in one assay. Our technique is based on multi-colour lentiviral self-inactivating (SIN) LeGO vector technology. RESULTS: We demonstrated the successful use of this approach for screening compounds against up to four HIV gag-pol variants (wild-type and three mutants) simultaneously. Importantly, the technique was adapted to Biosafety Level 1 conditions by utilising ecotropic pseudotypes. This allowed upscaling to a large-scale screening protocol exploited by pharmaceutical companies in a successful proof-of-concept experiment. CONCLUSIONS: The technology developed here facilitates fast screening for anti-HIV activity of individual agents from large compound libraries. Although drugs targeting gag-pol variants were used here, our approach permits screening compounds that target several different, key cellular and viral functions of the HIV life-cycle. The modular principle of the method also allows the easy exchange of various mutations in HIV sequences. In conclusion, the methodology presented here provides a valuable new approach for the identification of novel anti-HIV drugs.
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spelling pubmed-35601532013-02-04 A new system for parallel drug screening against multiple-resistant HIV mutants based on lentiviral self-inactivating (SIN) vectors and multi-colour analyses Prokofjeva, Maria M Riecken, Kristoffer Spirin, Pavel V Yanvarév, Dimitriy V Düsedau, Arne Ellinger, Bernhard Fehse, Boris Stocking, Carol Prassolov, Vladimir S AIDS Res Ther Methodology BACKGROUND: Despite progress in the development of combined antiretroviral therapies (cART), HIV infection remains a significant challenge for human health. Current problems of cART include multi-drug-resistant virus variants, long-term toxicity and enormous treatment costs. Therefore, the identification of novel effective drugs is urgently needed. METHODS: We developed a straightforward screening approach for simultaneously evaluating the sensitivity of multiple HIV gag-pol mutants to antiviral drugs in one assay. Our technique is based on multi-colour lentiviral self-inactivating (SIN) LeGO vector technology. RESULTS: We demonstrated the successful use of this approach for screening compounds against up to four HIV gag-pol variants (wild-type and three mutants) simultaneously. Importantly, the technique was adapted to Biosafety Level 1 conditions by utilising ecotropic pseudotypes. This allowed upscaling to a large-scale screening protocol exploited by pharmaceutical companies in a successful proof-of-concept experiment. CONCLUSIONS: The technology developed here facilitates fast screening for anti-HIV activity of individual agents from large compound libraries. Although drugs targeting gag-pol variants were used here, our approach permits screening compounds that target several different, key cellular and viral functions of the HIV life-cycle. The modular principle of the method also allows the easy exchange of various mutations in HIV sequences. In conclusion, the methodology presented here provides a valuable new approach for the identification of novel anti-HIV drugs. BioMed Central 2013-01-03 /pmc/articles/PMC3560153/ /pubmed/23286882 http://dx.doi.org/10.1186/1742-6405-10-1 Text en Copyright ©2013 Prokofjeva et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology
Prokofjeva, Maria M
Riecken, Kristoffer
Spirin, Pavel V
Yanvarév, Dimitriy V
Düsedau, Arne
Ellinger, Bernhard
Fehse, Boris
Stocking, Carol
Prassolov, Vladimir S
A new system for parallel drug screening against multiple-resistant HIV mutants based on lentiviral self-inactivating (SIN) vectors and multi-colour analyses
title A new system for parallel drug screening against multiple-resistant HIV mutants based on lentiviral self-inactivating (SIN) vectors and multi-colour analyses
title_full A new system for parallel drug screening against multiple-resistant HIV mutants based on lentiviral self-inactivating (SIN) vectors and multi-colour analyses
title_fullStr A new system for parallel drug screening against multiple-resistant HIV mutants based on lentiviral self-inactivating (SIN) vectors and multi-colour analyses
title_full_unstemmed A new system for parallel drug screening against multiple-resistant HIV mutants based on lentiviral self-inactivating (SIN) vectors and multi-colour analyses
title_short A new system for parallel drug screening against multiple-resistant HIV mutants based on lentiviral self-inactivating (SIN) vectors and multi-colour analyses
title_sort new system for parallel drug screening against multiple-resistant hiv mutants based on lentiviral self-inactivating (sin) vectors and multi-colour analyses
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3560153/
https://www.ncbi.nlm.nih.gov/pubmed/23286882
http://dx.doi.org/10.1186/1742-6405-10-1
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