title: GFAP promoter-driven RNA interference on TGF-β1 to treat liver fibrosis.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  GFAP promoter driven TGF-β1 pri-miRNA producing plasmids have the potential to be used for site-specific gene therapeutics to treat liver fibrosis.
authors.affiliation: Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, 19 South Manassas, Memphis, Tennessee 38103-3308, USA.
authors.forename: Ningning
authors.initials: N
authors.lastname: Yang
authors.forename: Ram I
authors.initials: RI
authors.lastname: Mahato
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D020868:Gene Silencing; D015316:Genetic Therapy / Q000379:methods; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics*; D055166:Hepatic Stellate Cells / Q000187:drug effects / Q000378:metabolism; D008103:Liver Cirrhosis / Q000235:genetics / Q000628:therapy*; D035683:MicroRNAs / Q000235:genetics; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors* / Q000235:genetics
chemical_list: D005904:Glial Fibrillary Acidic Protein; D035683:MicroRNAs; D034741:RNA, Small Interfering; D053773:Transforming Growth Factor beta1