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Partial promoter substitutions generating transcriptional sentinels of diverse signaling pathways in embryonic stem cells and mice.

Serup P, Gustavsen C, Klein T, Potter LA, Lin R, Mullapudi N, Wandzioch E, Hines A, Davis A, Bruun C, Engberg N, Petersen DR, Peterslund JM, Macdonald RJ, Grapin-Botton A, Magnuson MA, Zaret KS
Dis Model Mech. 2012 5 (6): 956-66

PMID: 22888097 · PMCID: PMC3484877 · DOI:10.1242/dmm.009696

Extracellular signals in development, physiology, homeostasis and disease often act by regulating transcription. Herein we describe a general method and specific resources for determining where and when such signaling occurs in live animals and for systematically comparing the timing and extent of different signals in different cellular contexts. We used recombinase-mediated cassette exchange (RMCE) to test the effect of successively deleting conserved genomic regions of the ubiquitously active Rosa26 promoter and substituting the deleted regions for regulatory sequences that respond to diverse extracellular signals. We thereby created an allelic series of embryonic stem cells and mice, each containing a signal-responsive sentinel with different fluorescent reporters that respond with sensitivity and specificity to retinoic acids, bone morphogenic proteins, activin A, Wnts or Notch, and that can be adapted to any pathway that acts via DNA elements.

MeSH Terms (24)

Activins Animals Base Sequence Bone Morphogenetic Proteins Embryo, Mammalian Embryonic Stem Cells Genetic Engineering Genetic Loci Humans Mice Molecular Sequence Data Mutation Promoter Regions, Genetic Proteins Rats Receptors, Notch Recombination, Genetic Response Elements RNA, Untranslated Sequence Deletion Signal Transduction Transcription, Genetic Tretinoin Wnt Signaling Pathway

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