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Publication : An ES cell system for rapid, spatial and temporal analysis of gene function in vitro and in vivo.

First Author  Mao J Year  2005
Journal  Nucleic Acids Res Volume  33
Issue  18 Pages  e155
PubMed ID  16221970 Mgi Jnum  J:165962
Mgi Id  MGI:4839160 Doi  10.1093/nar/gni146
Citation  Mao J, et al. (2005) An ES cell system for rapid, spatial and temporal analysis of gene function in vitro and in vivo. Nucleic Acids Res 33(18):e155
abstractText  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.
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