What activates Cre?
What activates Cre?
Inducible Cre activation is achieved using CreER (estrogen receptor) variant, which is only activated after delivery of tamoxifen. This is done through the fusion of a mutated ligand binding domain of the estrogen receptor to the Cre recombinase, resulting in Cre becoming specifically activated by tamoxifen.
How is Cre recombinase activated?
One of the ligand-dependent Cre recombinases is CreER recombinase, which consists of Cre fused to mutated hormone-binding domain of the estrogen receptor. The CreER recombinase is inactive but can be activated by the synthetic estrogen receptor ligand tamoxifen or 4-hydroxytamoxifen.
How does the Cre-lox system work?
General principle of Cre-loxP system Concerning the mechanism of Cre-loxP system, a single Cre recombinase recognizes two directly repeated loxP site, then the Cre excises the loxP flanked (floxed) DNA, thus creating two types of DNA with circular, excised and inactivated gene Y (Figure 1A).
What is a Cre activity?
Cre recombinase-mediated DNA recombination is an established method for conditional control of gene expression in animal models. Regulation of its activity has been accomplished to impart spatial and/or temporal control over recombination of the target gene.
How many base pairs is Cre?
34 base pair
This 34 base pair (bp) loxP recognition site consists of two 13 bp palindromic sequences which flank an 8bp spacer region….
| Cre recombinase | |
|---|---|
| Structure of a Cre recombinase enzyme (dimer) bound to its substrate DNA | |
| Identifiers | |
| Organism | Enterobacteria phage P1 |
| Symbol | cre |
How does tamoxifen induce Cre?
It utilizes a mutated estrogen receptor (ER) fused to Cre as a transgene (Cre-ER), which only becomes activated and then translocates into the nucleus upon binding of the active tamoxifen (TAM) metabolite 4-hydroxytamoxifen (4-OHT), which mutates the ER.
How does Cre ERT2 work?
The Cre-ERT2 fusion protein (Cre-ERT2) consists of Cre recombinase fused to a triple mutant form of the human estrogen receptor; which does not bind its natural ligand (17β-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT or tamoxifen) and, with …
What is the function of Cre recombinase?
Cre Recombinase is a Type I topoisomerase from bacteriophage P1 that catalyzes the site-specific recombination of DNA between loxP sites (1). The enzyme requires no energy cofactors and Cre-mediated recombination quickly reaches equilibrium between substrate and reaction products (2).
How does tamoxifen Cre work?
