B6.129S7-Kit^tm1Rosay/J

Stock number: 015813
Research areas: Research Tools

Description

This Kit^+/copGFP knock-in mutation strain expresses the copGFP variant of Green Fluorescent Protein (from the copepod Pontellina plumata) from the endogenous Kit locus. copGFP fluorescence mimics the endogenous Kit expression pattern. This mutant mouse strain may be useful for fluorescent visualization of KIT+ cells.

Detailed description

Mice that are heterozygous for this alelle are viable, normal in size and do not display any gross behavioral abnormalities. Heterozygotes exhibit white patches at the end of the tail, the belly and paws. Homozygotes exhibit an embryonic lethal phenotype. Mutant allele expression levels (mRNA) are similar to endogenous Kit expression in multiple tissues as detected by qPCR analysis (brain, heart, liver, lung, kidney, stomach, small intestine, colon). High expression levels are detected in lung. copGFP fluorescence is detected in KIT immuno-positive gastrointestinal interstitial cells of Cajal (ICC) by immunohistochemical and flow cytometric analysis.

When crossed with mice homozygous for the Lep^ob allele (see Stock No. 000632), loss in the intramuscular and myenteric populations of ICC in the intestine and colon can be visualized with confocal microscopy.

Development

The P1 artificial chromosome (PAC) clone 554-D1, containing exons 1-4 and a portion of intron 4 of the Kit gene, was modified by inserting copGFP (from the copepod Pontellina plumata), a Hprt-PGK cassette and the SV40 polyA signal, downstream of the start codon. The construct was electroporated into 129S7/SvEvBrd-Hprt^b-m2 derived AB2.2 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts. The resulting male chimeric animals were crossed to C57BL/6J female mice, and then backcrossed to the same for approximately 10 generations. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.

Allele

Symbol: Kit^tm1Rosay
Name: targeted mutation 1, Kenton M Sanders and Seungil Ro
MGI accession ID: MGI:4939904
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Kit
Marker name: KIT proto-oncogene receptor tyrosine kinase
Chromosome: 5
Strain of origin: 129S7/SvEvBrd-Hprt^b-m2
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: GFP fluorescence is detected in KIT immuno-positive gastrointestinal interstitial cells of Cajal (ICC) by immunohistochemical and flow cytometric analysis.
Molecular note: The P1 artificial chromosome (PAC) clone 554-D1, containing exons 1-4 and a portion of intron 4 of the Kit gene, was modified by insertion of copGFP (from the copepod Pontellina plumata), a Hprt-PGK cassette, and the SV40 polyA signal downstream of the start codon.