GFAP Promoter-Driven RNA Interference on TGF-β1 to Treat Liver Fibrosis
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The objective was to determine the role of promoters and miRNA backbone in shRNA-based hepatic stellate cell (HSC)-specific transforming growth factor (TGF)-β1 gene silencing. This is expected to avoid the side effect of non-specific TGF-β1 gene silencing.
Two most potent shRNAs targeting 769 and 1033 start sites of rat TGF-β1 mRNA were cloned into pSilencer 1.0 vector for enhanced TGF-β1 gene silencing. We then constructed HSC-specific pri-miRNA mimic and pri-miRNA cluster mimic expression plasmids in which shRNA expression was driven by a glial fibrillary acidic protein (GFAP) promoter to achieve HSC-specific TGF-β1 gene silencing to avoid nonspecific inhibition of TGF-β1 expression in other cells and organs.
These TGF-β1 pri-miRNA-producing plasmids showed the inhibition of proliferation and induced apoptosis of activated HSC-T6 cells. TGF-β1 pri-miRNA cluster mimic plasmids decreased TGF-β1 and collagen gene expression at both mRNA and protein levels.
GFAP promoter driven TGF-β1 pri-miRNA producing plasmids have the potential to be used for site-specific gene therapeutics to treat liver fibrosis.
KEY WORDSGFAP promoter liver fibrosis pri-miRNA mimics shRNA TGF-β1
This study was supported by a grant from the National Institutes of Health (to RIM, grant number: EB003922).
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