Dual-reporter system for specific tracing of insulin-producing cells

Abstract

[한글]

[영문]Islet transplantation offers a potential treatment for type 1 diabetes. Currently, islet graft survival can be assessed noninvasively only by measuring blood glucose, insulin, and C-peptide levels. However, these variables have limited value. To trace ?-cell survival or functional status, we constructed an adenovirus/adenoassociate virus hybrid vector (Hyb-DR) carrying the rat insulin II promoter-driven dual reporter (DR) genes with luciferase and green fluorescent protein (GFP) connected by the internal ribosome entry site. Luciferase activity increased and positive GFP expression was observed in ?-cell lines (MIN6N8 and INS-1E) infected with Hyb-DR. Using an in vivo imaging machine, the GFP signal was detected in the flank of nude mice 2 weeks after transplantation of Hyb-DR-infected MIN6N8 cells into the kidney capsule. Using non-?-cell lines (HepG2, FL83B, and YGIC5), coinfection of Hyb-DR with hybrid-carrying ?-cell-specific transcription factors also activated luciferase and GFP expression. This dual reporter system can provide quantitative and visual information about the functional islet mass and activation of the insulin gene. It may also be useful for ?-cell tracing in vivo