Lentiviral vectors with amplified β cell-specific gene expression
| Autor: | Dianne C. Skelton, Shundi Ge, Kit L. Shaw, Eszter Pais, Crooks Gm, Donald B. Kohn, Roger P. Hollis, Cinnamon L Hardee |
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| Rok vydání: | 2009 |
| Předmět: |
Transcriptional Activation
GAL4/UAS system Therapeutic gene modulation lineage-specificity gene amplification Genetic Vectors Green Fluorescent Proteins insulin promoter Gene Expression Biology Medical and Health Sciences Article Cell Line Promoter Regions Islets of Langerhans Mice 03 medical and health sciences 0302 clinical medicine Genetic Insulin-Secreting Cells Gene expression Genetics Animals Humans Insulin Promoter Regions Genetic Molecular Biology 030304 developmental biology Regulator gene 0303 health sciences Reporter gene Expression vector Activator (genetics) lentiviral vector Lentivirus Gene Transfer Techniques Biological Sciences Molecular biology Phosphoglycerate Kinase Organ Specificity 030220 oncology & carcinogenesis Molecular Medicine Heterologous expression Biotechnology |
| Zdroj: | Gene therapy Gene therapy, vol 16, iss 8 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/gt.2009.49 |
| Popis: | An important goal of gene therapy is to be able to deliver genes, so that they express in a pattern that recapitulates the expression of an endogenous cellular gene. Although tissue-specific promoters confer selectivity, in a vector-based system, their activity may be too weak to mediate detectable levels in gene-expression studies. We have used a two-step transcriptional amplification system to amplify gene expression from lentiviral vectors using the human insulin promoter. In this system, the human insulin promoter drives expression of a potent synthetic transcription activator (the yeast GAL4 DNA-binding domain fused to the activation domain of the Herpes simplex virus-1 VP16 activator), which in turn activates a GAL4-responsive promoter, driving the enhanced green fluorescent protein reporter gene. Vectors carrying the human insulin promoter did not express in non-beta-cell lines, but expressed in murine insulinoma cell lines, indicating that the human insulin promoter was capable of conferring cell specificity of expression. The insulin-amplifiable vector was able to amplify gene expression five to nine times over a standard insulin-promoter vector. In primary human islets, gene expression from the insulin-promoted vectors was coincident with insulin staining. These vectors will be useful in gene-expression studies that require a detectable signal and tissue specificity. |
| Databáze: | OpenAIRE |
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