C57BL/6-Tg(Ins2-luc/EGFP/TK)300Kauf/J

Stock number: 012943
Product name: B6.MIP-TF
Research areas: Diabetes and Obesity Research, Research Tools

Description

B6.MIP-TF mice harbor the tri-fusion transgene of three imaging reporters, luciferase/EGFP/thymidine kinase, under the control of mouse insulin promoter. Longitudinal noninvasive imaging of beta cells in the same transgenic animal by CCD and microPET, and the identification of beta cells at the cellular level by fluorescent microscopy is possible. This transgenic model provides a new tool for monitoring beta cells from the single cell level to non-invasive longitudinal assessments of beta cells in animal models of type 1 diabetes and type 2 diabetes.

Detailed description

C57BL/6J mice carrying tri-fusion transgene Ins2-luc/EGFP/Tk, commonly called MIP-TF, line 300 express a luciferase/enhanced green fluorescent protein/thymidine kinase fusion protein (luc/EGFP/Tk) under the control of mouse insulin promoter (Ins2) and are viable, fertile and exhibit no gross physical or behavioral abnormalities. Transgene expression, determined by luciferase enzymatic activity and green fluorescence is exclusive to the pancreas, specifically the insulin producing cells. Transgene expression appears to have no effect on weight, islet morphology, fasting blood glucose, or response to glucose challenge when compared to wild-type controls. This trifusion reporter model should enable longitudinal noninvasive imaging of beta cells in the same animal by cooled charge coupled device (CCD) and micro positron emission tomography (microPET), and identification of beta cells at the cellular level by fluorescent microscopy. There is a correlation between CCD and microPET signals from the pancreas region of the individual transgenic mice. After low dose streptozotocin treatment to induce type 1 diabetes, there is a reduction in the bioluminescence signals from the pancreas prior to the appearance of hyperglycemia. Conversely, pancreatic bioluminescence signals from the pancreas progressively increase (3.6 fold higher compared to control mice) in mice fed a high fat diet to model early type 2 diabetes. This transgenic model provides a new tool for monitoring beta cells from the single cell level to non-invasive longitudinal assessments of beta cells in animal models of type 1 diabetes and type 2 diabetes.

Development

The transgene consists of a mouse insulin promoter, (MIP, Ins2) driving a trifusion imagining reporter consisting of a bioluminescent reporter from a modified firefly luciferase gene (luc) fused to a fluorescent reporter, enhanced green fluorescent protein (EGFP) derived from the jellyfish Aequorea Victoria, which is fused to a positron emission tomography reporter, herpes simplex virus 1 sr39 thymidine kinase (HSV-1 sr39-TK). This Ins2-luc/EGFP/Tk transgene (commonly referred to as MIP-TF) was microinjected into C57BL/6 embryos. Several founders were identified and evaluated. The donating investigator reported that founder line 300 has been maintained on a C57BL/6 background (see SNP note below). In 2010, The T1DR received this transgenic strain and mated to C57BL/6J for 1 generation prior to sibling mating.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, all 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a C57BL/6N genetic background.

Allele

Symbol: Tg(Ins2-luc/EGFP/TK)300Kauf
Name: transgene insertion 300, Daniel Kaufman
MGI accession ID: MGI:4820879
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(Ins2-luc/EGFP/TK)300Kauf
Marker name: transgene insertion 300, Daniel Kaufman
Chromosome: UN
Strain of origin: C57BL/6
Expressed genes: HSV-TK,herpes simplex virus thymidine kinase,; luc,luciferase,firefly; GFP,Green Fluorescent Protein,
Site of expression: A luciferase/enhanced green florescent protein/thymidine kinase fusion protein (luc/EGFP/Tk) is expressed in insulin producing beta cells of the pancreas.
Molecular note: The transgene consists of a mouse insulin promoter, (MIP, Ins2) driving a trifusion imagining reporter consisting of a bioluminescent reporter from a modified firefly luciferase gene (luc) fused to a florescent reporter, enhanced green florescent protein (EGFP) derived from the jellyfish Aequorea Victoria, which is fused to a positron emission tomography reporter, herpes simplex virus 1 sr39 thymidine kinase (HSV-1 sr39-TK).