Recombinant LuIII Autonomous Parvovirus as a Transient Transducing Vector for Human Cells
| العنوان: | Recombinant LuIII Autonomous Parvovirus as a Transient Transducing Vector for Human Cells |
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| المؤلفون: | Jonathan O. Carlson, Françoise Maxwell, Solon L. Rhode, Ian H. Maxwell, Joe Corsini |
| المصدر: | Human Gene Therapy. 4:441-450 |
| بيانات النشر: | Mary Ann Liebert Inc, 1993. |
| سنة النشر: | 1993 |
| مصطلحات موضوعية: | biology, Parvovirus, viruses, Genetic Vectors, Clone (cell biology), biology.organism_classification, Genome, Virology, Molecular biology, Parvoviridae, law.invention, Blotting, Southern, Genes, Reporter, Transduction, Genetic, law, Genetics, Recombinant DNA, Humans, Molecular Medicine, Luciferase, Vector (molecular biology), Cloning, Molecular, Molecular Biology, HeLa Cells |
| الوصف: | Recombinants based on the genome of the autonomous parvovirus, LuIII, were constructed by replacing the viral coding sequences in an infectious clone (pGLu883) by a luciferase or beta-galactosidase reporter, which was linked to the viral P4 promoter. In cells cotransfected with either of these constructs, together with a plasmid supplying LuIII nonstructural and capsid proteins, excision and replication of the recombinant genome occurred. Transducing virions accumulated in the culture medium of the cotransfected cells, as assayed by reporter activity in recipient cells exposed to this medium. Transducing activity could be neutralized by antiserum to LuIII. Production of replicative form DNA and transducing virions were observed following cotransfection of HeLa, 293, or NB324K cells, in increasing order of efficiency. When homology existed between the recombinant genome and sequences flanking the viral genes in the helper construct, concomitant production of replication-competent, cytopathic virus was sometimes observed. This could be minimized by removal of the left end homology from the helper; by this means, preparations of luciferase transducing virus were obtained free from replication-competent virus. With such preparations, we observed luciferase expression (declining after 3 days) for up to 7 days in recipient HeLa cells. Hybridization of the recombinant viral DNA with strand-specific luciferase probes indicated packaging of both strands (as reported for LuIII), but with a several-fold excess of the (-) strand. We suggest that transducing-autonomous parvoviruses will be useful in gene transfer applications, possibly including gene therapy when only transient expression is desired. |
| تدمد: | 1557-7422 1043-0342 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a129fe9b88ddfc8f5b00914bbab879d7 https://doi.org/10.1089/hum.1993.4.4-441 |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....a129fe9b88ddfc8f5b00914bbab879d7 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15577422 10430342 |
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