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

• Published in: Gene therapy Vol. 16; no. 2; pp. 279 - 290
• Main Authors: Del Vecchio, C, Calistri, A, Lombardi, G, Celegato, M, Biasolo, M A, Palù, G, Parolin, C
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-02-2009; Nature Publishing Group
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Applied cell therapy and gene therapy; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Biotechnology; Cell Biology; Cell Line; Fundamental and applied biological sciences. Psychology; Gag protein; Gene Expression; Gene Therapy; Genetic aspects; Genetic vectors; Genetic Vectors - genetics; Health aspects; Health. Pharmaceutical industry; HIV; HIV (Viruses); HIV-1 - genetics; Human Genetics; Human immunodeficiency virus; Human immunodeficiency virus 1; Humans; Immune system; Industrial applications and implications. Economical aspects; Medical sciences; Nanotechnology; original-article; Packaging; Recombination, Genetic; Replicase; Replicon - genetics; Semliki Forest virus; Semliki forest virus - genetics; Trans-Activators - biosynthesis; Trans-Activators - genetics; Transcription; Transduction, Genetic; Transfection; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Viral proteins; Virion - genetics; Virions; Viruses; Italy; HIV-1 vector; recombinant virus particles; Semliki Forest virus; transduction; Semliki forest virus; Recombinant virus; Retroviridae; Togaviridae; Lentivirus; Virus; Particle; Production; Alphavirus; Transduction; Human immunodeficiency virus; Gene therapy; Vector
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/gt.2008.159

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/VSV
G
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.