Stable transduction of actively dividing cells via a novel adenoviral/episomal vector.
Many gene therapy indications would benefit from vectors capable of achieving efficient in vivo delivery and long-term transgene expression in either dividing or nondividing cells. Such vector systems are not yet available. To achieve both goals, we have used noncytotoxic E1- and E4-deleted adenoviral vectors as vehicles for delivering an Epstein-Barr virus-based self-replicating episome (replicon) via Cre/loxP site-specific recombination. Co-infection of human cells with a proreplicon-encoded and a Cre-expressing adenovirus resulted in efficient delivery and excision of a functional replicon in the absence of vector-induced cytotoxicity. In addition, replication and nuclear retention of the replicon in the cell progeny translated into a prolonged transgene expression in actively dividing cells, both in vitro and in vivo. Combining desired features from different viruses within a single hybrid vector system should expand the range of clinical indications currently amenable to gene transfer.
Leblois
H
H
Institut Gustave Roussy, CNRS-IGR-Rhône Poulenc Rorer UMR 1582, Villejuif, France. leblois@igr.fr
Roche
C
C
Di Falco
N
N
Orsini
C
C
Yeh
P
P
Perricaudet
M
M
D000260:Adenoviruses, Human / Q000235:genetics*; D001483:Base Sequence; D002455:Cell Division; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006367:HeLa Cells; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D019426:Integrases / Q000235:genetics; D007763:Lac Operon; D010957:Plasmids / Q000235:genetics*; D012093:Replicon / Q000235:genetics; D014161:Transduction, Genetic; D014162:Transfection; D014764:Viral Proteins
D017931:DNA Primers; D019309:Epstein-Barr Virus Nuclear Antigens; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases