Promoter Strength in Adenovirus Transducing Vectors: Down-Regulation of the Adenovirus E1A Promoter in 293 Cells Facilitates Vector Construction
| Autor: | David J. Orlicky, Blake Allen, Roderic L. Smith, Ian H. Maxwell, Jerome Schaack, Michael L. Bennett |
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| Rok vydání: | 2001 |
| Předmět: |
Gene Expression Regulation
Viral viruses Transgene Genetic Vectors Down-Regulation Transfection Cell Line Viral vector E1A promoter 03 medical and health sciences Transduction (genetics) 0302 clinical medicine Transduction Genetic Virology Humans Promoter Regions Genetic promoter strength transcription inhibition 030304 developmental biology Regulation of gene expression 0303 health sciences Rous sarcoma virus biology Adenoviruses Human HEK 293 cells Promoter biology.organism_classification Molecular biology adenovirus transducing vectors Adenovirus E1A Proteins 030217 neurology & neurosurgery HeLa Cells |
| Zdroj: | Virology. 291:101-109 |
| ISSN: | 0042-6822 |
| DOI: | 10.1006/viro.2001.1211 |
| Popis: | Most adenovirus transducing vectors have the cytomegalovirus major immediate-early (CMV) or the Rous sarcoma virus long terminal repeat (RSV) promoter driving expression of the transgene. Both of these promoters are highly active in transfection and transduction assays in 293 cells, in which transducing vectors are constructed and grown, and in HeLa cells. The CMV promoter exhibits rapid activation while the RSV promoter exhibits a lag prior to the onset of viral DNA replication in transduction assays. While the use of very strong promoters facilitates expression of the transgene, high-level expression of certain gene products hinders virus construction and growth. For such genes, the use of the adenovirus type 5 E1A promoter offers advantages. The E1A promoter exhibits modest activity in HeLa cells after transfection or transduction, but very little activity in 293 cells, suggesting that the E1A promoter would permit construction and growth of vectors encoding deleterious gene products that could not be constructed with the CMV and RSV promoters. This idea was tested through attempts to construct viruses encoding the immunoglobulin loop 6 and transmembrane regions of the prostaglandin F2α receptor regulatory protein (FPRP), a product that inhibits adenovirus vector construction for reasons that are not clear. Only the E1A promoter permitted construction and growth of the transducing vector encoding the fragment of FPRP. |
| Databáze: | OpenAIRE |
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