B6.Cg-Shh^{tm1(EGFP/cre)Cjt}/J

Stock number: 005622
Product name: Shhgfpcre
Research areas: Developmental Biology Research, Research Tools

Description

Fluorescence in these mice, consistent with the endogenous Shh locus, is detected in the distal posterior region of the limb buds of embryos aged embryonic day 10 to 12. Mice homozygous for the mutation develop a limited limb skeleton and lack digit 2. This mutant mouse strain may be useful in studies of limb patterning and development.

Detailed description

This strain expresses a fusion product involving Enhanced Green Fluorescent Protein (EGFP) and Cre recombinase from the endogenous Shh locus. EGFP and cre expression are consistent with the endogenous gene. Fluorescence is detected in the distal posterior region of the limb buds of embryos aged embryonic day 10 to 12 and colocalizes with the endogenous gene product (mRNA). The donating investigator reports that it is not uncommon for a mosaic expression pattern to be exhibited when the allele is inherited through the female germline. It is recommended that this allele be passed through the male germline when conducting experiments involving cre-induced recombination. Mice homozygous for the mutation develop a limited limb skeleton and lack digit 2. Homozygous mice are not viable or fertile. Heterozygous mutant mice are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities.

This mutant mouse strain may be useful in studies of limb patterning and development.

Development

A targeting vector containing a fusion product involving Enhanced Green Fluorescent Protein (EGFP) and Cre recombinase was inserted at the ATG of Shh and disrupted the sequence encoding the first 12 amino acids. The construct was electroporated into 129S6/SvEv-derived TC-1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to C57BL/6 mice, and the donating investigator reported that they were then backcrossed to the same for at least 5 generations.

Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664). The mice were then backcrossed to C57BL/6J (Stock No. 000664) for 8 generations (October 2018).

Allele

Symbol: Shh^{tm1(EGFP/cre)Cjt}
Name: targeted mutation 1, Clifford J Tabin
MGI accession ID: MGI:3053959
Generation method: Targeted
Attributes: Recombinase-expressing; Null/Knockout
Marker symbol: Shh
Marker name: sonic hedgehog
Chromosome: 5
Strain of origin: Not Specified
Expressed genes: cre,cre recombinase,bacteriophage P1; GFP,Green Fluorescent Protein,
Site of expression: Distal posterior region of the limb buds of embryos aged embryonic day 10 to 12; also a Cre strain.
Molecular note: A cassette containing an in frame fusion between GFP and cre was inserted at the ATG. Upon recombination, the first 35 base pairs after the ATG were deleted. Fluorescence was detected in the distal posterior region of E10-E12 embryos.