C57BL/6-Tg(Nkx3-1-EGFP*)18Zaw/J

Stock number: 039646
Product name: Nkx3.1(11)-d2EGFP
Research areas: Cancer Research, Cell Biology Research, Reproductive Biology Research, Research Tools

Description

Nkx3.1(11)-d2EGFP transgenic mice express a destabilized green fluorescent protein (d2GFP) under control of the the mouse Nkx3-1 promoter. These mice may be useful for tracking prostate epithelial cells and studying transcriptional regulation of the Nkx3-1 gene.

Detailed description

NK3 homeobox 1, encoded by Nkx3-1, is a transcription factor expressed in the prostate epithelium and acts as a negative regulator of epithelial cell growth in prostate tissue. Nkx3-1 mutations have been associated with prostate carcinogenesis.

These Nkx3.1(11)-d2EGFP mice harbor a transgene with a destabilized green fluorescent protein (d2GFP) sequence with an SV40 polyA sequence right after the start codon of the Nkx3-1. The d2EGFP has a half-life of ~2 hours.

Hemizygous and homozygous mice are viable and fertile with no overt phenotype. GFP expression is detected in the prostate starting around postnatal age P4. In adult mice, GFP is expressed in most luminal cells of the prostate epithelium and its expression recapitulates Nkx3-1 expression during androgen deprivation and re-administration. Fluorescence is measurable by flow cytometry, immunofluorescence and direct fluorescence. This Nkx3.1(11)-d2EGFP transgenic model may be useful for lineage tracking of prostate epithelial cells and studying transcriptional regulation of the Nkx3-1 gene.

Development

The Nkx3.1(11)-d2EGFP transgene was designed to express a destabilized green fluorescent protein (d2GFP) driven by the mouse NK3 homeobox 1 (Nkx3-1) promoter. This transgene construct is comprised of the 10.5-kb mouse genomic sequence (UCSC genome browser, mm9 Chr 14: 69808515-69819104) at the Nkx3-1 locus (-234/+10356) containing a 240-bp proximal promoter, exon 1, intron, exon 2, and ~6.5-kb 3' region as well as the d2EGFP (Clontech) and SV40 poly(A) sequences inserted right after the Nkx3-1 start codon. The construct (11671bp total length) was cloned into a pUC57 vector. Purified linearized DNA was microinjected into the pronuclei of fertilized C57BL/6N oocytes. Founder line 18 was established and maintained by the donating laboratory prior to sending males to The Jackson Laboratory Repository. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ (Stock No. 005304) oocytes.

Of note, the transgene integration site(s) and copy number have not been evaluated by the donating laboratory.

In 2024, 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 males sent to The Jackson Laboratory Repository. All 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating, including the rd8 recessive mutation associated with retinal degeneration on Chromosome 1 within the Crb1 locus. Additionally, 2 other markers were segregating for allele-type other than C57BL/6. Taken together, these data suggest the mice sent to The Jackson Laboratory Repository (and used for the frozen sperm in 2024) were on a predominantly C57BL/6 genetic background (mix of C57BL/6N;C57BL/6J) but also contained other undetermined genetic background contributions.

Allele

Symbol: Tg(Nkx3-1-EGFP*)18Zaw
Name: transgene insertion 18, Zhu A Wang
MGI accession ID: MGI:7660837
Generation method: Transgenic
Attributes: Reporter
Synonyms: Nkx3.1(11)-d2EGFP
Marker symbol: Tg(Nkx3-1-EGFP*)18Zaw
Marker name: transgene insertion 18, Zhu A Wang
Chromosome: UN
Strain of origin: C57BL/6N
Molecular note: The transgene was designed to express a destabilized green fluorescent protein (d2GFP) driven by the mouse NK3 homeobox 1 (Nkx3-1) promoter. Founder line 18 was established and maintained.