B6.Cg-Gt(ROSA)26Sor^{tm1(Sirt6)Mpgu}/J

Stock number: 040125
Product name: Tg.Sirt6
Strain synonyms: ROSA26-Sirt6-FLAG
Research areas: Cell Biology Research, Metabolism Research, Research Tools

Description

Tg.Sirt6 mice contain a Cre-inducible murine Sirt6 cDNA, with a C-terminal FLAG tag, inserted into the Gt(ROSA)26Sor locus. These mice are useful for studying effect of tissue-specific overexpression of SIRT6 in various pathophysiologic conditions.

Detailed description

Tg.Sirt6 mice contain a Cre-inducible knock-in of the mouse sirtuin 6 (Sirt6) cDNA with a C-terminal single FLAG tag introduced to the Gt(ROSA)26Sor locus. A floxed Neo/STOP cassette prevents FLAG-Sirt6 expression in the absence of Cre recombinase. After removal of the loxP-flanked STOP cassette, FLAG-SIRT6 is produced in Cre-expressing cells and daughter cells.

Sirt6 encodes a lysine deacylase of histones and non-histone proteins. It is implicated in the regulation of many key cellular processes such as metbolism homeostasis, immunity, inflammation, and genomic stability. Tg.Sirt6 mice are viable and fertile. Breeding females with male Pgk1-Cre mice (Stock No. 020811), results in whole-body FLAG-Sirt6-overexpressing mice. These mice are useful for studying effect of tissue-specific overexpression of SIRT6 in various pathophysiologic conditions.

Development

The Tg.Sirt6 targeting vector was designed with (from 5' to 3') a splice acceptor site, a loxP-flanked cassette containing a neomycin resistance gene (neo) and two polyadenylation signals, and a single FLAG-tagged mouse sirtuin 6 (Sirt6) cDNA sequence followed by a polyadenylation signal. This targeting vector also included 5' and 3' arms of homology for site-directed recombination into the Gt(ROSA)26Sor locus. The construct was electroporated into (C57BL/6NTac x 129S6/SvEvTac)F1-derived iTL BA1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and resulting Tg.Sirt6 mice were bred to C57BL/6J mice (Stock No. 000664) for at least 20 generations by the donating laboratory. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.



In 2025, 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. Two of the 43 markers throughout the genome were segregating with 129. Also, 3 of the 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. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Gt(ROSA)26Sor^{tm1(Sirt6)Mpgu}
Name: targeted mutation 1, Mahesh P Gupta
MGI accession ID: MGI:7525638
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Inserted expressed sequence; Epitope tag
Synonyms: ROSA26-Sirt6-FLAG; Sk.T6Tg; Tg.Sirt6
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Marker synonyms: beta geo; Gtrgeo26; Gtrosa26; R26; ROSA26; SETD5-AS1; Thumpd3as1
Chromosome: 6
Strain of origin: (C57BL/6NTac x 129S6/SvEvTac)F1
Molecular note: The targeting vector was designed to insert the Sirt6-FLAG cDNA into the Gt(ROSA)26Sor driven by its promoter and controlled by a STOP cassette. The STOP cassette in the vector consisted of a splice acceptor and the floxed (PGK)/gb2neoPGKpolyA2xSV40pA sequence and was inserted in intron 1 of ROSA26 locus. Mouse Sirt6 cDNA with a C-terminal FLAG tag was cloned immediately downstream of the STOP cassette. The Sirt6-FLAG sequence was followed by a BGHpA bovine growth hormone polyadenylation (BGHpA) sequence.