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RRE-deleting self-inactivating and self-activating HIV-1 vectors for improved safety

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Author(s): Narasimhachar Srinivasakumar

Journal: PeerJ
ISSN 2167-8359

Volume: 1;
Start page: e84;
Date: 2013;
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Keywords: RRE | Gene therapy | Vector | Lentivirus | HIV | HIV-1 vector | Self-inactivating | Self-activating

ABSTRACT
Retroviruses have been shown to efficiently delete sequences between repeats as a consequence of the template switching ability of the viral reverse transcriptase. To evaluate this approach for deriving safety-modified lentiviral vectors, we created HIV-1 vectors engineered to delete the Rev-response element (RRE) during reverse-transcription by sandwiching the RRE between two non-functional hygromycin phosphotransferase sequences. Deletion of the RRE during reverse-transcription lead to the reconstitution of a functional hygromycin phosphotransferase gene in the target cell. The efficiency of functional reconstitution, depending on vector configuration, was between 12% and 23%. Real-time quantitative PCR of genomic DNA of cells transduced with the RRE-deleting vectors that were selected using an independent drug resistance marker, which measured both functional and nonfunctional recombination events, indicated that the overall efficiency of RRE deletion of hygromycin phosphotransferase gene, was between 73.6% and 83.5%.
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Tango Jona
Tangokurs Rapperswil-Jona