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 PMID:22888097  

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

Palle Serup | Carsten Gustavsen | Tino Klein | Leah A Potter | Robert Lin | Nandita Mullapudi | Ewa Wandzioch | Angela Hines | Ashley Davis | Christine Bruun | Nina Engberg | Dorthe R Petersen | Janny M L Peterslund | Raymond J Macdonald | Anne Grapin-Botton | Mark A Magnuson | Kenneth S Zaret
Disease models & mechanisms | 2012

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.

Pubmed ID: 22888097

Associated grants

  • Agency: NIDDK NIH HHS, United States
    Id: U01 DK072473
  • Agency: NIDDK NIH HHS, United States
    Id: U01 DK072503
  • Agency: NIDDK NIH HHS, United States
    Id: U19 DK072495

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This is a list of tools and resources that we have found mentioned in this publication.


PipMaker and MultiPipMaker (tool)

RRID:SCR_011806

PipMaker computes alignments of similar regions in two DNA sequences. Moreover, MultiPipMaker can be requested to compute a true multiple alignment of the input sequences and return a nucleotide-level view of the results.

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Rosa26(228.AR8.TA.mCherry)Mgn (Gt(ROSA)26Sortm2.1(mix1b-mCherry)Mgn) (tool)

RRID:BCBC_4153

This mouse line was generated by RMCE in the Rosa26[LCA] allele. In this mouse line, the Rosa26 gene sequence (-228 to +81) was replaced by the Foxh1/Smad2-dependent, Mix2-derived Activin response element (AR8) fused to a TATA-mCherry reporter. These mice may be useful to assess Activin, Nodal, and TGFbeta responsiveness.

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C57BL/6J (tool)

RRID:IMSR_JAX:000664

Mus musculus with name C57BL/6J from IMSR.

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Mutant Mouse Resource and Research Center (biomaterial supply resource)

RRID:SCR_002953

National public repository system for mutant mice. Archives and distributes scientifically valuable spontaneous and induced mutant mouse strains and ES cell lines for use by biomedical research community. Includes breeding/distribution facilities and information coordinating center. Mice strains are cryopreserved, unless live colony must be established. Live mice are supplied from production colony, from colony recovered from cryopreservation, or via micro-injection of cell line into host blastocysts. MMRRC member facilities also develop technologies to improve handling of mutant mice, including advances in assisted reproductive techniques, cryobiology, genetic analysis, phenotyping and infectious disease diagnostics.

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