What is tamoxifen inducible Cre?
Introduction. Tamoxifen (TAM)-inducible Cre/loxP is one of the most widely used inducible systems for gene regulation. This is in part due to the fact that it enables gene control both spatially and temporally.
How long does tamoxifen last in mice?
Our results revealed that tamoxifen and its metabolites were completely degraded within 8 days in young adult C57BL/6J mice, while the age-matched FVB/NJ male mice displayed more effective degradation. Moreover, aged C57BL/6J mice were unable to metabolize all substances within 8 days.
What is CAG Cre?
STOCK Tg(CAG-cre/Esr1*)5Amc/J These CAGGCre-ERTM mice express a fusion protein consisting of Cre recombinase joined to the ligand-binding domain of a modified mouse estrogen receptor and can be used to generate tamoxifen-induced, Cre-mediated targeted deletions when bred with strains carrying a floxed gene.
How do you inject tamoxifen in mice?
To inject tamoxifen by oral gavage, use a 1 mL syringe and a 22-gauge feeding needle (100 μL to inject 1 mg). Place the mouse with its belly on a surface and restrain at its neck, tightly enough to keep its head immobilized but also to allow proper breathing.
What does tamoxifen do to mice?
Tamoxifen is widely used for the induction of genomic recombination in mice (double-)transgenic for floxed genes and Tamoxifen specific estrogen receptors (ER) coupled to Cre-recombinase (supplemental Table 1) (Hayashi and McMahon 2002).
Is tamoxifen toxic to mice?
Ways to deliver Tamoxifen to Mice Tamoxifen is a hazardous material – make sure to handle it properly. It is also light sensitive, therefore making fresh solutions is recommended.
Can tamoxifen be dissolved in water?
All Answers (14) Tamoxifen is water insoluble and could be dissolved in methanol, ethanol, propanol, propylene glycol, DMSO, or chloroform. However, these solvents are toxic, so many studies used peanut oil, or corn oil (shaking overhight 37°C!) to dissolve it.
How does tamoxifen induce gene expression?
A double injection of 0.15 mg/g body weight tamoxifen induces GFP expression in epithelial cells and fiber cells. Single injections or lower doses of tamoxifen are less effective in inducing GFP expression.
How to induce recombination in mice with tamoxifen?
For adult mice, a standard dose of 100µl tamoxifen/corn oil solution (above) is effective for inducing recombination. Administer tamoxifen via intraperitoneal injection (using an ACUC approved injection procedure) once every 24 hours for a total of 5 consecutive days. As a safeguard, sanitize injection site with 70% ethanol prior to injection.
What is the wait period for tamoxifen in mice?
For Cre characterization work at the Jackson Laboratory, there is a 7‐day waiting period between the final injection and necropsy/histological analysis. Throughout the course of tamoxifen injections and any post‐injection wait period, mice should be closely monitored for any adverse reactions to the treatment.
How do you administer tamoxifen in ACUC?
Administer tamoxifen via intraperitoneal injection (using an ACUC approved injection procedure) once every 24 hours for a total of 5 consecutive days. As a safeguard, sanitize injection site with 70% ethanol prior to injection.
What is the best way to store tamoxifen?
Madisen, L., et. al., Nat Neurosci. 2010; 13 (1): 133–140 overnight at 37°C. Tamoxifen is light sensitive, and should be made and stored in a light‐ blocking vessel (amber, or foil wrapped).