Efficient Recombination in Diverse Tissues by a Tamoxifen-Inducible Form of Cre: A Tool for Temporally Regulated Gene Activation/Inactivation in the Mouse
| Autor: | Andrew P. McMahon, Shigemi Hayashi |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Genetically modified mouse
Transcriptional Activation gene activation Transgene Recombinant Fusion Proteins 4-hydroxy-tamoxifen Mutagenesis (molecular biology technique) Mice Transgenic Biology Nervous System Gene Expression Regulation Enzymologic Embryonic and Fetal Development Mice Viral Proteins Pregnancy inducible Animals Site-specific recombinase technology inactivation Molecular Biology mouse Crosses Genetic Regulation of gene expression Recombination Genetic Reporter gene Integrases Gene Expression Regulation Developmental Cre Cell Biology Molecular biology Embryonic stem cell Fusion protein Tamoxifen Amino Acid Substitution Receptors Estrogen Enzyme Induction Mutagenesis Site-Directed Female Cre-ERTM estrogen receptor bacteriophage P1 Developmental Biology |
| Zdroj: | Developmental Biology. 244(2):305-318 |
| ISSN: | 0012-1606 |
| DOI: | 10.1006/dbio.2002.0597 |
| Popis: | In recent years, the Cre integrase from bacteriophage P1 has become an essential tool for conditional gene activation and/or inactivation in mouse. In an earlier report, we described a fusion protein between Cre and a mutated form of the ligand binding domain of the estrogen receptor (Cre-ERTM) that renders Cre activity tamoxifen (TM) inducible, allowing for conditional modification of gene activity in the mammalian neural tube in utero. In the current work, we have generated a transgenic mouse line in which Cre-ERTM is ubiquitously expressed to permit temporally regulated Cre-mediated recombination in diverse tissues of the mouse at embryonic and adult stages. We demonstrate that a single, intraperitoneal injection of TM into a pregnant mouse at 8.5 days postcoitum leads to detectable recombination in the developing embryo within 6 h of injection and efficient recombination of a reporter gene in derivatives of all three germ layers within 24 h of injection. In addition, by varying the dose of TM injected, the percentage of cells undergoing a recombination event in the embryo can be controlled. Dose-dependent excision induced by TM was also possible in diverse tissues in the adult mouse, including the central nervous system, and in cultured cells derived from the transgenic mouse line. This inducible Cre system will be a broadly useful tool to modulate gene activity in mouse embryos, adults, and culture systems where temporal control is an important consideration. |
| Databáze: | OpenAIRE |
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