An ES cell system for rapid, spatial and temporal analysis of gene function in vitro and in vivo
| Autor: | Jeffery R. Barrow, Joe E. Vaughan, Andrew P. McMahon, Junhao Mao, Jill A. McMahon |
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| Rok vydání: | 2005 |
| Předmět: |
RNA
Untranslated Time Factors Cellular differentiation Biology Cell Line Receptors G-Protein-Coupled Mice Viral Proteins 03 medical and health sciences 0302 clinical medicine Genetics Animals Cell Lineage Hedgehog Proteins Gene Alleles 030304 developmental biology Neurons Regulation of gene expression 0303 health sciences Integrases Chimera Stem Cells Proteins Cell Differentiation Embryo Mammalian Smoothened Receptor Embryonic stem cell Molecular biology Hedgehog signaling pathway Cell biology Tamoxifen Gene Expression Regulation Doxycycline Trans-Activators Methods Online Stem cell Smoothened 030217 neurology & neurosurgery Signal Transduction |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gni146 |
| Popis: | We describe a versatile genetic system for rapid analysis of mammalian gene function. In this, loss of reporter activity in a novel embryonic stem (ES) cell line enables rapid identification of targeting to the ubiquitously expressed Rosa26 locus. Subsequent regulation of gene activity is governed by a dual regulatory strategy utilizing two drugs, Tamoxifen and Doxycycline. To illustrate this approach, a dominant allele of Smoothened was introduced into this cell line, enabling regulated activation of Hedgehog signaling. By coupling Cre-loxP dependent activation with tetracycline dependent transcription in a single allele, we established a conditional method to control Smoothened activity and neural progenitor specification in differentiating ES cells in vitro and in chimeric embryos in vivo When crossed to an appropriate Cre driver strain, gene activity can also be temporally regulated within a specific cell lineage. This platform will facilitate rapid analysis of gene function in the mouse. |
| Databáze: | OpenAIRE |
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