Coupling of glucose response element from L-type pyruvate kinase and G6Pase promoter enhances glucose responsive activity in hepatoma cells.
Type 1 diabetes results from the autoimmune destruction of pancreatic beta-cells, which leads to severe insulin deficiency. Insulin gene therapy provides an attractive approach to cure diabetes. The critical factor for insulin gene therapy in surrogate cells is to select an appropriate site for insulin expression and a tissue-specific promoter that is responsive to both physiological glucose and insulin concentrations. A novel chimeric promoter, (GIRE)n-G6Pase, consisting of a 1.6 kb glucose 6-phosphatase (G6Pase) promoter and a segment of the regulatory element derived from the L-type pyruvate kinase (L-PK) promoter, was designed to provide strong and tight control of insulin expression in liver. One or three copies of GIRE were linked to the G6Pase promoter, which showed a stronger promoter activity than the G6Pase promoter alone. The chimeric promoter was inhibited by insulin in a dosage-dependent manner and activated by glucose, two features essential for glucose metabolism. The promoter activity is conserved between species and highly specific for liver cells. The construction of a chimeric promoter with stronger and more sensitive responsive activity to glucose and insulin in liver cells could further advance studies in insulin gene therapy.
Lan
Michael S
MS
The Research Institute for Children, Departments of Pediatrics and Genetics, Children's Hospital, Louisiana State University Health Sciences Center, 200 Henry Clay Avenue, New Orleans, LA 70118, USA. mlan@chnola-research.org
Wang
Hong-Wei
HW
Chong
James
J
Breslin
Mary B
MB
D000818:Animals; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000235:genetics*; D005947:Glucose / Q000494:pharmacology*; D005952:Glucose-6-Phosphatase / Q000235:genetics*; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D009928:Organ Specificity / Q000187:drug effects; D011770:Pyruvate Kinase / Q000235:genetics*; D051381:Rats; D020218:Response Elements / Q000235:genetics*
D007328:Insulin; D011770:Pyruvate Kinase; D005952:Glucose-6-Phosphatase; D005947:Glucose