A novel, helper-dependent, adenovirus-retrovirus hybrid vector: stable transduction by a two-stage mechanism.
We have developed a novel vector system that uses a helper-dependent adenoviral vector as a carrier to deliver a fully functional retrovirus vector. The helper-dependent adenovirus (HDAd) can accommodate large inserts, provide high titers, and infect nondividing as well as dividing cells. However, adenoviral DNA is rarely integrated into the host cell genome, and its episomal expression is transient. Therefore we inserted a replication-competent, ecotropic retrovirus vector containing the green fluorescent protein (GFP) reporter gene as a second-stage component. The well-characterized host species tropism of each vector component provided a stringent biological assay system that demonstrates the two-stage transduction mechanism of the hybrid vector, because the adenovirus stage can efficiently transduce human cells but cannot replicate in murine cells, and conversely, the ecotropic retrovirus stage cannot enter human cells but can efficiently proliferate in murine cells, resulting in permanent integration and progressive spread of reporter gene expression.
Soifer
Harris
H
Institute for Genetic Medicine, University of Southern California, 2250 Alcazar Street, CSC-240, Los Angeles, California 90033, USA.
Higo
Collin
C
Logg
Christopher R
CR
Jih
Lily Ja-Lu
LJ
Shichinohe
Toshiaki
T
Harboe-Schmidt
Erik
E
Mitani
Kohnosuke
K
Kasahara
Noriyuki
N
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015139:Blotting, Southern; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins; D006801:Humans; D019426:Integrases; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics*; D010957:Plasmids; D011995:Recombination, Genetic; D012190:Retroviridae / Q000235:genetics*; D020079:Terminal Repeat Sequences; D014161:Transduction, Genetic; D019076:Transgenes; D014407:Tumor Cells, Cultured; D014764:Viral Proteins; D016662:Virus Integration
D008164:Luminescent Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases