Functional silencing of hepatic microsomal glucose-6-phosphatase gene expression in vivo by adenovirus-mediated delivery of short hairpin RNA.
An expression cassette containing mouse U6 polymerase III promoter directing expression of short hairpin RNA (shRNA) targeting murine microsomal glucose-6-phosphatase (G6P) transcript was generated. This construct was packaged into an adenoviral (AdV) backbone and viral stocks generated. Mice injected intravenously with AdV-G6PshRNA exhibited a significant reduction in postprandial glucose levels and had significantly elevated steady-state hepatic glycogen stores. Target gene silencing was confirmed by measurements demonstrating a significant reduction in both hepatic G6P transcript level and phosphohydrolase activity. These findings provide evidence that AdV delivery of expressed shRNA can be a productive tool to explore gene function in vivo.
Huang
Alan
A
Millennium Pharmaceuticals, 75 Sidney St., Cambridge, MA 02139, USA.
Chen
Yan
Y
Wang
Xinzhong
X
Zhao
Shanchuan
S
Su
Nancy
N
White
David W
DW
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D005952:Glucose-6-Phosphatase / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D006801:Humans; D007739:L Cells; D008112:Liver Glycogen / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs; D008862:Microsomes, Liver / Q000201:enzymology*; D051381:Rats
D001786:Blood Glucose; D008112:Liver Glycogen; D008164:Luminescent Proteins; D035683:MicroRNAs; D049452:Green Fluorescent Proteins; D005952:Glucose-6-Phosphatase