Lentiviral vectors with amplified β cell-specific gene expression View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2009-08

AUTHORS

Kit L. Shaw, Eszter Pais, Shundi Ge, Cinnamon Hardee, Dianne Skelton, Roger Hollis, Gay M. Crooks, Donald B. Kohn

ABSTRACT

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. More... »

PAGES

gt200949

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/gt.2009.49

DOI

http://dx.doi.org/10.1038/gt.2009.49

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1027274281

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/19440227


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