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Analysis of human immunodeficiency virus type 1 vector cis- and trans-acting elements production by means of Semliki Forest virus

Gene Therapy volume 16, pages 279–290 (2009)Cite this article

Abstract

Recombinant Semliki Forest virus (SFV) is an attractive viral vector system owing to its ability to allow high efficiency of viral protein expression. To produce recombinant pseudotyped human immunodeficiency virus type 1 (HIV-1) virions, we designed a chimeric SFV/HIV vector system that contains both the HIV-1 cis- and trans-acting elements under the transcriptional control of the SFV replicase and investigated the ability of the hybrid SFV/HIV system to produce lentiviral particles capable of transducing target cells. Co-transfection of target cells with the two helper SFV packaging system RNAs along with each SFV/Gag-Pol, SFV/VSVG as well as SFV/HIV-1 vector unit replicon led to the generation of efficient transducing competent recombinant SFV/HIV particles. In contrast, co-transduction of target cells with the SFV/HIV chimeric virions produced recombinant particles with low transducing ability. Our data suggest that both the genomic and the subgenomic RNAs containing the HIV-1 vector unit were negatively selected for incorporation into recombinant particles, despite the fact that the SFV-driven HIV-1 vector replicon was the only one containing a lentiviral packaging sequence. The results of this study provide insights relevant to the design of chimeric lentiviral vectors.

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Acknowledgements

This work was supported by grants from Regione Veneto, MIUR (PRIN-2005), Istituto Superiore di Sanità (Rome-AIDS Project nos. 30G.24 and 30G.55) and University of Padova (Ex 60% and ATENEO 2006) to AC, GP and CP. MC is a PhD student in the Virology and Microbial Biotechnologies program at the University of Padova. We thank F Salandin for artwork.

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  1. C Del Vecchio and A Calistri: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Histology, Microbiology and Medical Biotechnologies, University of Padova, Padova, Italy

    C Del Vecchio, A Calistri, G Lombardi, M Celegato, M A Biasolo & G Palù

  2. Department of Biology, University of Padova, Padova, Italy

    C Parolin

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  1. C Del Vecchio
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  2. A Calistri
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Correspondence to C Parolin.

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This paper is dedicated to the memory of R Parolin.

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Del Vecchio, C., Calistri, A., Lombardi, G. et al. Analysis of human immunodeficiency virus type 1 vector cis- and trans-acting elements production by means of Semliki Forest virus. Gene Ther 16, 279–290 (2009). https://doi.org/10.1038/gt.2008.159

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