B6.Cg-Tg(Ins1-HIST1H2BB/mCherry)5091Mhsg/J

Stock number: 028589
Product name: mIns1-H2b-mCherry
Research areas: Diabetes and Obesity Research, Research Tools

Description

mIns1-H2b-mCherry mice emit nuclear-localized mCherry fluorescence in all pancreatic beta cells. Mice are useful for visualization and FACS isolation of pancreatic beta cells, allowing for studies of beta-cell specific gene expression.

Detailed description

mIns1-H2b-mCherry transgenic mice express an H2B-mCherry fusion protein (coding sequence for the human HIST1H2BB gene [histone 1 H2bb] followed C-terminally by a monomeric red fluorescent protein sequence) under the control of the murine insulin I (Ins1) promoter. Insulin is expressed only be the beta cells of pancreatic islets. Mice homozygous for the transgene are viable and fertile, and are normoglycemic with no sign of glucose intolerance. mIns1-H2b-mCherry mice emit nuclear-localized mCherry fluorescence in all pancreatic beta cells. Fluorescence is not seen in any other cells. Mice are useful for visualization and FACS isolation of pancreatic beta cells, allowing for studies of beta-cell specific gene expression. A similar line of mice, Stock No. 006866, show a more mosaic expression pattern of DsRed, with less than 1% of beta cells labeled. Stock No. 006864, also expressing EGFP in pancreatic beta cells, has been shown to only express the fluorophore in 50% of insulin-positive cells.

Development

The mIns1-H2b-mCherry transgene was designed with the murine insulin I (Ins1) promoter driving expression of an H2B-mCherry fusion protein (coding sequence for the human HIST1H2BB gene [histone 1 H2bb] with a C-terminal monomeric red fluorescent protein sequence). The transgene was microinjected into CB6F1 fertilized oocytes, and mice from founder line number 5091 were bred to C57BL/6NHsd mice to establish a colony. The founder was determined to transmit the transgene to approximately 75% of F1 offspring, indicative of 2 independent transgene insertion sites. One of these sites was subsequently determined to be silent and was bred out, resulting in mIns1-H2b-mCherry colony that transmits at the expected Mendelian frequency. Mice from this line were backcrossed to C57BL/6NHsd mice for 13 generations. These mice were also bred to other reporter mice and Cre recombinase expressing mice on C57BL/6 backgrounds during their development. All incidental genetic contributions have been bred out of the colony. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).

Of note, these mice do not contain a human growth factor cassette(hGH).

Allele

Symbol: Tg(Ins1-HIST1H2BB/mCherry)5091Mhsg
Name: transgene insertion 5091, Mark Huising
MGI accession ID: MGI:5698910
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(Ins1-HIST1H2BB/mCherry)5091Mhsg
Marker name: transgene insertion 5091, Mark Huising
Chromosome: UN
Strain of origin: (BALB/c x C57BL/6)F1
Expressed genes: H2BC3,H2B clustered histone 3,human; RFP,Red Fluorescent Protein,coral
Molecular note: The transgene was designed with the murine insulin I (Ins1) promoter driving expression of an H2B-mCherry fusion protein (coding sequence for the human HIST1H2BB gene [histone 1 H2bb] with a C-terminal monomeric red fluorescent protein sequence). The founder was determined to transmit the transgene to approximately 75% of F1 offspring, indicative of 2 independent transgene insertion sites. One of these sites was subsequently determined to be silent and was bred out. Nuclear-localized mCherry fluorescence is observed in all pancreatic beta cells. Fluorescence is not seen in any other cells.