Selection of potent non-toxic inhibitory sequences from a randomized HIV-1 specific lentiviral short hairpin RNA library.

RNA interference (RNAi) has been considered as an efficient therapeutic approach against the human immunodeficiency virus type 1 (HIV-1). However, to establish a durable inhibition of HIV-1, multiple effective short hairpin RNAs (shRNAs) need to be stably expressed to prevent the emergence of viral...

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Autores principales:	Carola Pongratz, Benjamin Yazdanpanah, Hamid Kashkar, Maik J Lehmann, Hans-Georg Kräusslich, Martin Krönke
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/dc8f010013af4309bf754b005fde4b6f
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spelling	oai:doaj.org-article:dc8f010013af4309bf754b005fde4b6f2021-11-18T07:03:34ZSelection of potent non-toxic inhibitory sequences from a randomized HIV-1 specific lentiviral short hairpin RNA library.1932-620310.1371/journal.pone.0013172https://doaj.org/article/dc8f010013af4309bf754b005fde4b6f2010-10-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20949027/?tool=EBIhttps://doaj.org/toc/1932-6203RNA interference (RNAi) has been considered as an efficient therapeutic approach against the human immunodeficiency virus type 1 (HIV-1). However, to establish a durable inhibition of HIV-1, multiple effective short hairpin RNAs (shRNAs) need to be stably expressed to prevent the emergence of viral escape variants. In this study, we engineered a randomized lentiviral H1-promoter driven shRNA-library against the viral genome. Potent HIV-1 specific shRNAs were selected by ganciclovir treatment of cell lines stably expressing the cDNA of Herpes Simplex Virus thymidine kinase (HSV-TK) fused to HIV-1 nucleotide sequences. More than 50% of 200 selected shRNAs inhibited an HIV-1 based luciferase reporter assay by more than 70%. Stable expression of some of those shRNAs in an HIV-1 permissive HeLa cell line inhibited infection of wild-type HIV-1 by more than 90%. The combination of a randomized shRNA-library directed against HIV-1 with a live cell selection procedure yielded non-toxic and highly efficient HIV-1 specific inhibitory sequences that could serve as valuable candidates for gene therapy studies.Carola PongratzBenjamin YazdanpanahHamid KashkarMaik J LehmannHans-Georg KräusslichMartin KrönkePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 10, p e13172 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Carola Pongratz Benjamin Yazdanpanah Hamid Kashkar Maik J Lehmann Hans-Georg Kräusslich Martin Krönke Selection of potent non-toxic inhibitory sequences from a randomized HIV-1 specific lentiviral short hairpin RNA library.
description	RNA interference (RNAi) has been considered as an efficient therapeutic approach against the human immunodeficiency virus type 1 (HIV-1). However, to establish a durable inhibition of HIV-1, multiple effective short hairpin RNAs (shRNAs) need to be stably expressed to prevent the emergence of viral escape variants. In this study, we engineered a randomized lentiviral H1-promoter driven shRNA-library against the viral genome. Potent HIV-1 specific shRNAs were selected by ganciclovir treatment of cell lines stably expressing the cDNA of Herpes Simplex Virus thymidine kinase (HSV-TK) fused to HIV-1 nucleotide sequences. More than 50% of 200 selected shRNAs inhibited an HIV-1 based luciferase reporter assay by more than 70%. Stable expression of some of those shRNAs in an HIV-1 permissive HeLa cell line inhibited infection of wild-type HIV-1 by more than 90%. The combination of a randomized shRNA-library directed against HIV-1 with a live cell selection procedure yielded non-toxic and highly efficient HIV-1 specific inhibitory sequences that could serve as valuable candidates for gene therapy studies.
format	article
author	Carola Pongratz Benjamin Yazdanpanah Hamid Kashkar Maik J Lehmann Hans-Georg Kräusslich Martin Krönke
author_facet	Carola Pongratz Benjamin Yazdanpanah Hamid Kashkar Maik J Lehmann Hans-Georg Kräusslich Martin Krönke
author_sort	Carola Pongratz
title	Selection of potent non-toxic inhibitory sequences from a randomized HIV-1 specific lentiviral short hairpin RNA library.
title_short	Selection of potent non-toxic inhibitory sequences from a randomized HIV-1 specific lentiviral short hairpin RNA library.
title_full	Selection of potent non-toxic inhibitory sequences from a randomized HIV-1 specific lentiviral short hairpin RNA library.
title_fullStr	Selection of potent non-toxic inhibitory sequences from a randomized HIV-1 specific lentiviral short hairpin RNA library.
title_full_unstemmed	Selection of potent non-toxic inhibitory sequences from a randomized HIV-1 specific lentiviral short hairpin RNA library.
title_sort	selection of potent non-toxic inhibitory sequences from a randomized hiv-1 specific lentiviral short hairpin rna library.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/dc8f010013af4309bf754b005fde4b6f
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Selection of potent non-toxic inhibitory sequences from a randomized HIV-1 specific lentiviral short hairpin RNA library.

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