Tamoxifen induced cre

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    Tamoxifen induced cre


    , a fusion protein between a mutated ligand binding domain of the human estrogen receptor (ER) and the Cre recombinase, the activity of which can be induced by 4-hydroxy-tamoxifen (OHT), but not natural ER ligands. We have recently characterized a new ligand-dependent recombinase, Cre-ER efficiency of these two ligand-inducible recombinases to generate temporally-controlled somatic mutations, we have engineered transgenic mice expressing a Lox P-flanked (floxed) transgene reporter and either Cre-ER under the control of the bovine keratin 5 promoter that is specifically active in the epidermis basal cell layer. No background recombinase activity could be detected, while recombination was induced in basal keratinocytes upon OHT administration. Interestingly, a dose-response study showed that Cre-ER The efficient introduction of somatic mutations in a given gene, at a given time, in a specific cell type, will greatly facilitate studies of gene function in vertebrates. We have previously established a temporally-controlled site-specific recombination system in mice using a conditional Cre/lox system. The fusion of the Cre recombinase with a mutated ligand binding domain (LBD) of the human estrogen receptor (ER) resulted in a tamoxifen-dependent Cre recombinase, Cre-ER in transgenic mice could be used for the generation of site-specific somatic mutations in a spatio-temporally-controlled manner. Short-term tamoxifen treatments have low acute toxicity and cause no severe abnormalities in mice (3). Cre-Lox recombination is a site-specific recombinase technology, used to carry out deletions, insertions, translocations and inversions at specific sites in the DNA of cells. It allows the DNA modification to be targeted to a specific cell type or be triggered by a specific external stimulus. It is implemented both in eukaryotic and prokaryotic systems. The Cre-lox recombination system has been particularly useful to help neuroscientists to study the brain in which complex cell types and neural circuits come together to generate cognition and behaviors. NIH Blueprint for Neuroscience Research has created several hundreds of Cre driver mouse lines which are currently used by the worldwide neuroscience community. The system consists of a single enzyme, Cre recombinase, that recombines a pair of short target sequences called the Lox sequences. This system can be implemented without inserting any extra supporting proteins or sequences.

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    Nov 6, 2009. Addition of tamoxifen induces a conformational change in the ER LBD and release from Hsp90 which induces nuclear translocation and Cre. Cre-Lox recombination is a site-specific recombinase technology, used to carry out deletions. it is possible for translocation events to be catalysed by Cre induced recombination. Once both sequences of the mutated Cre and the loxP genes are located in one mouse, the absence of tamoxifen will result in the shuttling. Refined protocols of tamoxifen injection for inducible DNA. DNA recombination in astrocytes, based on the GLAST-CreERT2/loxP system.

    Adult neurogenesis is a tightly regulated process continuously taking place in the central nervous system of most mammalian species. doi: 10.1073/pnas. Pub Med Abstract | Cross Ref Full Text | Google Scholar Packard, M. In neuroscience research, transgenic animals bearing the tamoxifen-inducible Cre ER activates the expression of YFP in multipotent neural stem cells upon tamoxifen application. Humoral factors, such as the levels of estrogens, have been reported to affect the hippocampal neurogenesis. The application of tamoxifen, a mixed agonist/antagonist of the estrogen receptor that permeates the blood-brain-barrier, could thus influence adult neurogenesis. Although the functions of adult neurogenesis are yet to be fully deciphered, a reciprocal interaction between rates of neurogenesis on the one hand and learning and mood regulation on the other hand, has been suggested. The impact of tamoxifen on neurogenesis and behavior was therefore addressed following five daily applications according to the open field test, the elevated plus maze, and Morris water maze. In addition, the impact of short-term tamoxifen application on progenitor cell proliferation, morphology, and fate in the neurogenic niche of the dentate gyrus were investigated. Tissue-specific and time-dependent control of in vivo gene disruption may be achieved using conditional knockout strategies in transgenic mice. Fusion of mutant estrogen receptor ligand-binding domains to Cre recombinase (Cre-ER, Mer Cre Mer) combined with cardiac-directed gene expression has been used to generate several cardiac-specific tamoxifen-inducible Cre-expressing mouse lines. Such mice have successfully been used to generate Cre-lox P-mediated gene disruption in an inducible manner in the myocardium in vivo. However, information is sparse regarding the tamoxifen dosage, the time course of gene disruption and whether different administration routes differ in efficiency in obtaining gene disruption in the myocardium. We have evaluated these parameters in We thank Carsten Lund for advice on feed, Dag Markus Eide, National Institute of Public Health, for lending us mouse feeders, Roy Trondsen for designing mouse feeders, Marianne Lunde Sneve for technical assistance, Heidi Kvaløy and Ulla H. Enger for help with testing tamoxifen feed pellets. KBA was funded by Southeastern Norway Regional Health Authority and University of Oslo EMBIO senior fellow grants.

    Tamoxifen induced cre

    Tamoxifen-Induced Cre-loxP Recombination Is Prolonged in., Cre-Lox recombination - Wikipedia

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  6. Mar 19, 2002. Tamoxifen-Inducible Form of Cre A Tool for. Temporally Regulated Gene Activation/Inactivation in the Mouse. Shigemi Hayashi and Andrew P.

    • Efficient Recombination in Diverse Tissues by a Tamoxifen-Inducible..
    • Refined protocols of tamoxifen injection for inducible DNA. - Nature.
    • Inducible Cre mice. - NCBI.

    We have produced Cre transgenic mice in which excision is tamoxifen inducible and occurs in a widespread mosaic pattern. We utilized our Cre excision. Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis comparison of the recombinase activity of the tamoxifen-inducible Cre-ERT. Aug 2, 2007. with the tamoxifen-inducible fusion protein consisting of Cre recombinase. To induce recombination of conditional alleles in neurons of the.

     
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