C57BL/6J (BL6), B6;129-Sirt1tm1Ygu/J (Sirt1f/f), and C57BL/6J–Tg(Itgax-cre,-EGFP)4097Ach/J (CD11c–Cre-GFP) mice were purchased at 6–7 weeks of age from The Jackson Laboratory (Bar Harbor, ME). Sirt1f/f mice, in which two loxP sites flank Sirt1 exon 4, were crossed to CD11c–Cre-GFP transgene mice. As the Sirt1f/f mice were on a mixed C57BL/6J;129 background, we backcrossed the Sirt1f/f-CD11c–Cre progeny to a C57BL/6J background for 6 generations. Deletion of exon 4 produces a truncated protein that lacks catalytic activity, causing a Sirt1-null genotype (24 (link)). Thus, Cre+ mice lack a functional SIRT1 in CD11chigh cells. Sirt1f/f-CD11c–Cre mouse breeding took place in-house at the University of Michigan (Ann Arbor, MI). All work involving animals was reviewed and approved by the University of Michigan University Committee on Care and Use of Animals.
B6 129 sirt1tm1ygu j
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B6;129-Sirt1tm1Ygu/J is a transgenic mouse strain. It expresses a targeted mutation in the Sirt1 gene.
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4 protocols using b6 129 sirt1tm1ygu j
1
SIRT1 Knockout in CD11c+ Cells
2
Sirt1 Knockout in CD11c+ Cells
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C57BL/6J (BL6), B6;129-Sirt1tm1Ygu/J(Sirt1f/f), and C57BL/6J-Tg (Itgax-Cre-EGFP) 4097Ach/J (CD11c-Cre-GFP) mice were purchased at 6–7 weeks of age from the Jackson Laboratory (Bar Harbor, ME). Sirt1f/f mice, in which two loxP sites flank Sirt1 exon 4, were crossed to CD11c-Cre-GFP transgenic mice. As the Sirt1f/f mice were on a mixed C57BL/6J;129 background, we backcrossed the Sirt1f/f-CD11c-Cre progeny to a C57BL/6J background for six generations. Deletion of exon 4 produces an internally truncated protein that lacks catalytic activity, causing a Sirt1-null genotype [64 ]. Thus, Cre+ mice lack a functional SIRT1 in CD11chigh cells. Sirt1f/f-CD11c-Cre (SIRT1-/-) mouse breeding took place in-house at the University of Michigan (Ann Arbor, MI).
3
Generating Myeloid-Specific SIRT1 Knockout Mice
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SIRT1loxP/loxP mice (B6.129-Sirt1tm1Ygu/J) and LysM-Cre mice (B6.129P2-lyz2tm1(cre)Ifo/J) were purchased from Jackson Laboratory (Bar Harbor, ME, USA). SirT1loxP/loxP and LysM-Cre mice were crossed to obtain mSIRT1 KO mice. To avoid potential variations that could be contributed by gender and genetic background, male mice from the F2 generation, SIRT1loxP/loxPLysM-Cre+/+ (mSIRT1 KO) and SIRT1loxP/loxPLysM-Cre−/− (WT), were used for the studies. All of the experimental animals used in this study were maintained under the protocol approved by the Institutional Animal Care and Use Committee of the Gyeongsang National University (GLA-100917-M0092).
4
Tamoxifen-Inducible SIRT1 Knockout and Overexpression in Mice
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All experimental procedures performed were approved by the Institutional Animal Care and Use Committee at Washington University and conformed to the National Institutes of Health Guidelines for the Care and Use of Animals in Research. We obtained 12‐ to 14‐week‐old male C57BL/6J mice from Jackson Laboratories (Bar Harbor, ME). Whole body adult‐inducible SIRT1‐knockout and SIRT1‐overexpressing mice were generated as described previously.24 In brief, mice with a targeted Sirt1 floxed mutant allele (B6;129‐Sirt1tm1Ygu/J) were obtained from Jackson Laboratories and bred with mice expressing whole‐body tamoxifen‐inducible Cre (B6.129‐Gt[ROSA]26Sortm1[cre/ERT2]Tyj/J) obtained from Jackson Laboratories to generate tamoxifen‐inducible whole‐body SIRT1‐knockout mice and wild‐type littermate controls. Hemizygous constitutive whole‐body SIRT1‐overexpressing mice (B6. CgTg[Sirt1]ASrn/J) and noncarriers on the same background were also obtained from Jackson Laboratories and bred to generate constitutive whole‐body SIRT1‐overexpressing mice (Sirt1‐Tg) and littermate controls. All animals were acclimated under a 12‐hour light/dark cycle controlled environment with free access to water and a standard mouse chow diet.
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