B6.129S-Atoh1^{tm5.1(Cre/PGR)Hzo}/J

Stock number: 013594
Research areas: Developmental Biology Research, Neurobiology Research, Research Tools

Description

This knockin mutation replaces the coding sequence of the (Atoh1) gene with a modified Cre-recombinase-progesterone receptor fusion protein (Cre-PR). The fusion gene activity is inducible by administration of RU486.

Detailed description

A targeting vector was designed to replace the coding sequence of the atonal homolog 1 (Atoh1) gene with a modified Cre-recombinase-progesterone receptor fusion protein (Cre-PR), abolishing gene function. Homozygous Math1^Cre*PR mice die before birth due to central respiratory failure. Heterozygous mice are viable,fertile, and normal in size. The Math1^Cre*PR/+ allele has expression of the Cre*PR fusion protein under control of the Math1 promoter/enhancer elements. Cre*PR fusion gene activity is inducible; observed only 6-12 hours after administration of RU486, a competitive progesterone receptor antagonist. As such, when Math1^Cre*PR/+ mice are bred with mice containing loxP-flanked sequence, RU486-inducible Cre-mediated recombination will result in deletion of the floxed sequences in the Math1-expressing cells of the offspring. As such Math1 is expressed in the hindbrain, specifically in the conscious proprioceptive nuclei of the cortex at E10.5, and in most unconscious proprioceptive lineages in the cerebellum at E12.5-E14.5. These mutant mice are useful for the studying the characterization and time of formation of Math1-expressing cell lineages, as well as genetic and developmental links between proprioception, interoception, hearing, and arousal.

Development

A targeting vector was designed to insert a modified Cre-recombinase-progesterone receptor fusion protein (Cre-PR) downstream of the start codon of the atonal homolog 1 (Atoh1) gene, followed by a frt-flanked neomycin (neo) selection cassette. This construct was electroporated into 129S7/SvEvBrd-Hprt1^b-m2-derived AB2.2 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric mice were bred with 129S4/SvJaeSor-Gt(ROSA)26Sor^tm1(FLP1)Dym mutant mice to remove the neo cassette. The resulting Math1^Cre*PR/+ mice were subsequently mated with inbred C57BL/6 mice for at least 8 generations. Upon arrival at The Jackson Laboratory, these mice were bred with C57BL/6J mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Atoh1^{tm5.1(cre/PGR)Hzo}
Name: targeted mutation 5.1, Huda Y Zoghbi
MGI accession ID: MGI:4418334
Generation method: Targeted
Attributes: Recombinase-expressing; Inducible
Marker symbol: Atoh1
Marker name: atonal bHLH transcription factor 1
Chromosome: 6
Strain of origin: 129S7/SvEvBrd-Hprt^b-m2
Expressed genes: cre,cre recombinase,bacteriophage P1
Site of expression: RU486-inducible Cre-mediated recombination will result in deletion of Math1 in the hindbrain, specifically in the conscious proprioceptive nuclei of the cortex at E10.5, and in most unconscious proprioceptive lineages in the cerebellum at E12.5-E14.5.
Molecular note: All coding sequence was replaced by a modified cre/progesterone receptor fusion construct followed by an frt flanked PGK-neomycin selection cassette. The selection cassette was subsequently excised by crossing to mice carrying Gt(ROSA)26Sor^tm1(FLP1)Dym. In the resulting mice, the inducible cre is under the control of the endogenous gene promoter.