STOCK Gt(ROSA)26Sor^{tm1(Neurod1-EGFP)Able}/BfriJ

Stock number: 034697
Product name: ^LSLNeurod1
Research areas: Cell Biology Research, Endocrine Deficiency Research, Research Tools

Description

^LSL-Neurod1 mice contain Cre-inducible Neurod1-EGFP from the Gt(ROSA)26Sor locus. These mice are a useful tool when studying endocrine research and neurology.

Detailed description

^LSL-Neurod1 mice contain from 5' to 3', a floxed-STOP cassette containing a neomycin (neo) resistance gene, full-length mouse neurogenic differentiation 1 (Neurod1) cDNA, an internal ribosome entry site (IRES), and an enhanced green fluorescence protein (EGFP). This targeting vector was inserted into the Gt(ROSA)26Sor locus. Neurod1 is a transcription factor required for enteroendocrine progenitor differentiation and maturation. Mice that are homozygous for the targeted insertion are viable and fertile.

In the absence of Cre, Neurod1 gene expression is prevented by the STOP cassette. After removal of the loxP-flanked neo cassette via cre-mediated recombination, Neurod1 and EGFP are expressed, allowing for labeling cells/tissues where Cre recombinase is expressed.

When bred to Villin-cre mice, expressing cre recombinase in the small intestine, the resulting offspring have a 4.4 fold increase in the expression of Neurod1 in intestinal cells. They exhibit an increase in the number of enteroendocrine cells (EECs), and EE specific transcription factors and hormones. There is also a substantial reduction of Paneth cell marker expression, although the expressions of enterocyte-, tuft- and goblet-cell specific markers are largely not affected.

In these mice, EGFP can be detected by immunofluorescent microscopy and immunostaining.

Development

The ^LSL-Neurod1 targeting vector was designed with, from 5' to 3', a floxed-STOP cassette containing a neomycin (neo) resistance gene, full-length mouse neurogenic differentiation 1 (Neurod1) cDNA, an internal ribosome entry site (IRES), and an enhanced green fluorescence protein (EGFP). This targeting vector was inserted into the Gt(ROSA)26Sor locus. The construct was electroporated into C57BL/6N-derived JM8.F6 embryonic stem (ES) cells and correctly targeted ES cells were injected into albino B6(Cg)-Tyr^c-2J/J blastocysts (Stock No. 000058). The resulting chimeric animals were crossed to B6(Cg)-Tyr^c-2J/J mice for at least 20 generations by the donating lab (see SNP note below). The Tyr^c-2J allele was bred out of this colony. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

In 2020, a 48 SNP (single nucleotide polymorphism) panel analysis, with 43 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. Markers across the genome were segregating with known and unknown markers, suggesting an incomplete backcross. Also, 3 of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating.

Allele

Symbol: Gt(ROSA)26Sor^{tm1(Neurod1-EGFP)Able}
Name: targeted mutation 1, Andrew B Leiter
MGI accession ID: MGI:6384644
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter
Synonyms: ^LSLNeurod1; Rosa26^{loxp-STOP-loxP}Neurod1-IRES-eGFP
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Marker synonyms: beta geo; Gtrgeo26; Gtrosa26; R26; ROSA26; SETD5-AS1; Thumpd3as1
Chromosome: 6
Strain of origin: C57BL/6N
Site of expression: Enteroendocrine progenitor cells.
Molecular note: The targeting vector was designed with, from 5' to 3', a floxed-STOP cassette containing a neomycin (neo) resistance gene, full-length mouse neurogenic differentiation 1 (Neurod1) cDNA, an internal ribosome entry site (IRES), and an enhanced green fluorescence protein (EGFP). This targeting vector was inserted into the Gt(ROSA)26Sor locus.