Recombinant AAV-2 harboring gfp-antisense/ribozyme fusion sequences monitor transduction, gene expression, and show anti-HIV-1 efficacy

• Published in: Gene therapy Vol. 6; no. 7; pp. 1231 - 1238
• Main Authors: HÖRSTER, A, TEICHMANN, B, HORMES, R, GRIMM, D, KLEINSCHMIDT, J, SCZAKIEL, G
• Format: Journal Article
• Language: English
• Published: Basingstoke Nature Publishing Group 01.07.1999
• Subjects: Acquired Immunodeficiency Syndrome - therapy; Adeno-associated virus 2; Antiviral agents; Biological and medical sciences; Biotechnology; CD4-Positive T-Lymphocytes; Cell fusion; Cell Line; Dependovirus - genetics; Fundamental and applied biological sciences. Psychology; Fusion protein; Gene Expression; Gene therapy; Genetic Therapy - methods; Genetic Vectors - genetics; Green Fluorescent Proteins; Health. Pharmaceutical industry; HIV; HIV-1 - genetics; Human immunodeficiency virus; Human immunodeficiency virus 1; Humans; Industrial applications and implications. Economical aspects; Luciferases - genetics; Luminescent Proteins - genetics; Replication; RNA, Antisense; RNA, Catalytic - genetics; Toxicity; Transfection; Virus Replication; Human; Immunopathology; Adenoviridae; Ribozyme; HIV-1 virus; Retroviridae; AIDS; Gene expression; Immune deficiency; Lentivirus; Infection; Virus; Associated virus; Cell line; Viral disease; Antiviral; Human immunodeficiency virus; Gene therapy; Hybrid gene
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/sj.gt.3300955

Vector-mediated delivery of potentially antivirally active genes is a key step in somatic gene therapy including therapeutic approaches against AIDS. A crucial technical prerequisite is to monitor DNA transfer into target cells. Here, we describe recombinant infectious particles derived from the adeno-associated virus type 2 (AAV-2) that are suitable to deliver effective HIV-1-directed antisense and ribozyme genes into target cells. To monitor transduction, we designed and tested a number of fusions between indicator-coding sequences of luciferase or gfp with effective HIV-1-directed antisense or ribozyme sequences. The combination of an indicator function and an antiviral func- tion in cis allows successful identification of transduced cells and measurement of effects on the replication of HIV-1 in antisense/ribozyme-expressing cells only. The fusion genes were shown to express the indicator genes. Inhibition of HIV-1 replication mediated by the antisense/ribozyme portion of the fusion transcripts was similar to parental constructs and neither acute nor long-term toxicity of fusion genes and their gene products was observed. These results suggest the use of rAAV constructs described here as tools to study the transducibility of target cells, gene expression and efficacy of HIV-1-directed antisense and ribozyme genes.