| DOI | Resolve DOI: https://doi.org/10.1007/s12033-016-9987-1 |
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| Author | Search for: Robert, Marc-André1 ; Search for: Lytvyn, Viktoria1 ; Search for: Deforet, Francis1 ; Search for: Gilbert, Rénald1 ; Search for: Gaillet, Bruno |
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| Name affiliation | - National Research Council Canada. Human Health Therapeutics
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| Format | Text |
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| Type | Article |
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| Journal title | Molecular Biotechnology |
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| ISSN | 1073-6085
1559-0305 |
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| Subject | virus-like particles; HIV-1 Gag; VLP production; protein delivery; protein engineering; green fluorescent protein; transcription factor |
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| Abstract | Virus-like particles (VLPs) derived from retroviruses and lentiviruses can be used to deliver recombinant proteins without the fear of causing insertional mutagenesis to the host cell genome. In this study we evaluate the potential of an inducible lentiviral vector packaging cell line for VLP production. The Gag gene from HIV-1 was fused to a gene encoding a selected protein and it was transfected into the packaging cells. Three proteins served as model: the green fluorescent protein and two transcription factors—the cumate transactivator (cTA) of the inducible CR5 promoter and the human Krüppel-like factor 4 (KLF4). The sizes of the VLPs were 120–150 nm in diameter and they were resistant to freeze/thaw cycles. Protein delivery by the VLPs reached up to 100% efficacy in human cells and was well tolerated. Gag-cTA triggered up to 1100-fold gene activation of the reporter gene in comparison to the negative control. Protein engineering was required to detect Gag-KLF4 activity. Thus, insertion of the VP16 transactivation domain increased the activity of the VLPs by eightfold. An additional 2.4-fold enhancement was obtained by inserting nuclear export signal. In conclusion, our platform produced VLPs capable of efficient protein transfer, and it was shown that protein engineering can be used to improve the activity of the delivered proteins as well as VLP production. |
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| Publication date | 2016-11-09 |
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| Language | English |
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| Peer reviewed | Yes |
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| NPARC number | 23001201 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction |
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Record identifier | 33088f30-02da-47a8-8f42-3384097211bf | | Record created | 2017-01-05 |
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| Record modified | 2017-01-05 |
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