These floxed mutant mice possess loxP sites flanking exon 6 of the targeted Rptor (regulatory associated protein of MTOR, complex 1) gene. This strain may be useful for studying mTOR-dependent regulation of ketogenesis and other cellular processes in response to feeding and fasting.
David Sabatini, Whitehead Institute
These floxed mutant mice possess loxP sites flanking exon 6 of the targeted Rptor (regulatory associated protein of MTOR, complex 1) gene. Mice that are homozygous for this allele are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. When these mutant mice are bred to C57BL6-Tg(AlbCre)21Mgn (Stock No. 003574) mice, which direct liver-specific expression of Cre, mice exhibit a decrease in liver size yet produce ketones for hours after feeding. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 6 deleted in the cre-expressing tissues. This strain may be useful for studying mTOR (mammalian or mechanistic target of rapamycin)-dependent regulation of ketogenesis and other cellular processes in response to feeding and fasting.
A targeting vector was designed to insert a loxP site upstream of exon 6 followed by a frt-flanked neomycin (neo) resistance cassette, followed by loxP sites flanking exon 6 of the regulatory associated protein of MTOR (Mechanistic target of rapamycin), complex 1 (Rptor) gene. The construct was electroporated into 129/SvEv-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into BALB/c blastocysts and resulting chimeric mice were bred to C57BL/6 mice. Heterozygous offspring were bred with Flp transgenic mice on a 129 background to delete the neo cassette, and progeny were crossed to remove the Flp-expressing transgene, resulting in a colony homozygous for the floxed-Rptor (Rptorflox/flox) allele. The donating investigator reports that these mice were backcrossed to C57BL/6 (see SNP note below) at least 4 generations. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 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. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 1 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.
|Allele Name||targeted mutation 1.1, David M Sabatini|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Allele Synonym(s)||RaploxP; Raptorf|
|Gene Symbol and Name||Rptor, regulatory associated protein of MTOR, complex 1|
|Strain of Origin||129S/SvEv|
|Molecular Note||Exon 6 was flanked by 5' and 3' LoxP sites located 111 bp upstream and 547 bp downstream, respectively, and an FRT flanked neo cassette was inserted via homologous recombination. The neo cassette was removed by flp mediated recombination.|
When maintaining a live colony, homozygous mice may be bred together.
When using the B6.Cg-Rptortm1.1Dmsa/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #013188 in your Materials and Methods section.
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