Stable integration of transgenes delivered by a retrotransposon-adenovirus hybrid vector.
Helper-dependent adenoviruses show great promise as gene delivery vectors. However, because they do not integrate into the host chromosome, transgene expression cannot be maintained indefinitely. To overcome these limitations, we have inserted an L1 retrotransposon/transgene element into a helper-dependent adenovirus to create a novel chimeric gene delivery vector. Efficient adenovirus-mediated delivery of the L1 element into cultured human cells results in subsequent retrotransposition and stable integration of the transgene. L1 retrotransposition frequency was found to correlate with increasing multiplicity of infection by the chimeric vector, and further retrotransposition from newly integrated elements was not observed on prolonged culture. Therefore, this vector, which utilizes components of low immunogenic potential, represents a novel two-stage gene delivery system capable of achieving high titers via the initial helper-dependent adenovirus stage and permanent transgene integration via the retrotransposition stage.
Soifer
H
H
Institute for Genetic Medicine, University of Southern California, Los Angeles, CA 90033, USA.
Higo
C
C
Kazazian
H H
HH
Moran
J V
JV
Mitani
K
K
Kasahara
N
N
D000256:Adenoviridae / Q000235:genetics*; D015139:Blotting, Southern; D002478:Cells, Cultured; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006378:Helper Viruses / Q000235:genetics*; D006801:Humans; D020084:Long Interspersed Nucleotide Elements / Q000235:genetics*; D008164:Luminescent Proteins / Q000378:metabolism; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D018626:Retroelements / Q000235:genetics*; D013223:Statistics as Topic; D014161:Transduction, Genetic; D019076:Transgenes / Q000235:genetics*; D014407:Tumor Cells, Cultured
D005819:Genetic Markers; D008164:Luminescent Proteins; D018626:Retroelements; D049452:Green Fluorescent Proteins