Universität zu Köln

Pongratz, Carola, Yazdanpanah, Benjamin, Kashkar, Hamid, Lehmann, Maik J., Kraeusslich, Hans-Georg and Kroenke, Martin (2010). Selection of Potent Non-Toxic Inhibitory Sequences from a Randomized HIV-1 Specific Lentiviral Short Hairpin RNA Library. PLoS One, 5 (10). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203

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Abstract

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 wildtype 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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Pongratz, Carola UNSPECIFIED UNSPECIFIED UNSPECIFIED Yazdanpanah, Benjamin UNSPECIFIED UNSPECIFIED UNSPECIFIED Kashkar, Hamid UNSPECIFIED UNSPECIFIED UNSPECIFIED Lehmann, Maik J. UNSPECIFIED UNSPECIFIED UNSPECIFIED Kraeusslich, Hans-Georg UNSPECIFIED UNSPECIFIED UNSPECIFIED Kroenke, Martin UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-494415
DOI:	10.1371/journal.pone.0013172
Journal or Publication Title:	PLoS One
Volume:	5
Number:	10
Date:	2010
Publisher:	PUBLIC LIBRARY SCIENCE
Place of Publication:	SAN FRANCISCO
ISSN:	1932-6203
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language IMMUNODEFICIENCY-VIRUS TYPE-1; SMALL INTERFERING RNAS; MAMMALIAN-CELLS; PHAGE POLYMERASE; GENE-THERAPY; SIRNA; EXPRESSION; INFECTION; SHRNA; ESCAPE Multiple languages Multidisciplinary Sciences Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/49441