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

ABSTRACT 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

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material as well as M Dittmar and H-G Kräusslich for supporting HIV-1 inhibition studies. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Deutsches Krebsforschungszentrum,

Forschungsschwerpunkt Angewandte Tumorvirologie, Im Neuenheimer Feld 242, Heidelberg, 69120, Germany A Hörster, B Teichmann, R Hormes, D Grimm, J Kleinschmidt & G Sczakiel Authors * A

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gfp-antisense/ribozyme fusion sequences monitor transduction, gene expression, and show anti-HIV-1 efficacy. _Gene Ther_ 6, 1231–1238 (1999). https://doi.org/10.1038/sj.gt.3300955 Download

citation * Received: 31 December 1998 * Accepted: 28 March 1999 * Published: 09 August 1999 * Issue Date: July 1999 * DOI: https://doi.org/10.1038/sj.gt.3300955 SHARE THIS ARTICLE Anyone you

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Springer Nature SharedIt content-sharing initiative KEYWORDS * adeno-associated virus type 2 (AAV-2) * antisense RNA, HIV-1-directed * CD34, transduction by rAAV * green fluorescent protein

(gfp) * human immunodeficiency virus type 1 (HIV-1) * ribozyme, HIV-1-directed