Overview

Mice harboring the pR26-228 AR8-TA-mCherry reporter express mCherry in cells and tissues that receive SMAD2/3 mediated signaling induced by Nodal/Activin. This strain allows live monitoring of SMAD2/3-mediated signaling events seen in embryonic development as well as during regeneration and homeostasis in adults.

Donating Investigator

Mark A. Magnuson, Vanderbilt University School of Medicine

Genetic overview

Genetic Background	Generation
000664 C57BL/6J

Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn}

Allele Type	Gene Symbol	Gene Name
Targeted	Gt(ROSA)26Sor	gene trap ROSA 26, Philippe Soriano

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Research Applications

Research Tools

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Details

Detailed Description

Mice harboring the pR26-228 AR8-TA-mCherry reporter are viable and fertile, with expression of mCherry under the control of the activin response element from the Xenopus laevis mix1b (Mix.2) promoter. The red fluorescent protein, mCherry, is expressed in cells and tissues that receive SMAD2/3 mediated signaling induced by Nodal/Activin. The Nodal signal transduction pathway is involved in mesoendoderm induction, patterning of the nervous system, and determination of dorsal- ventral axis in vertebrate embryos.
This strain allows live monitoring of
SMAD2/3-mediated signaling events seen in particular lineages and
stages in embryonic development as well as during regeneration and
homeostasis in adults. Additionally, it allows FACS-based isolation of
responding cells.

Development

Using the ROSA26^LCA or Loxed Cassette Acceptor allele (Gt(ROSA)26Sor ^tm1Mgn), which contains a loxP-flanked (lox71, lox2272) selection cassette in the ROSA26 promoter, a donor cassette is inserted into the acceptor allele via recombinase mediated cassette exchange (RMCE). The donor cassette or signaling sentinel construct includes a frt-flanked pgk-hygromycin and an activin response element (AR8) from the Xenopus laevis mix1b reporter fused to a TATA-mCherry reporter, which replaced the ROSA26 promoter sequence (-4 kb to -228 bp).

The construct was electroporated into 129S6/SvEvTac-derived TL1 ES cells. Correctly targeted ES cells were injected into blastocysts from C57BL/6J mice. Offspring were bred with B6;SJL-Tg(ACTFLPe)9205Dym/J transgenic mice to delete the pgk-hygromycin cassette. The donating investigator reported that the mice were backcrossed to C57BL/6J mice for 7 generations prior to sending to MMRRC at JAX.

Expression Data

Expressed Gene	RFP, Red Fluorescent Protein, coral
Site of Expression	Red fluorescent protein, mCherry, is expressed in cells and tissues involved in the Nodal signal transduction pathway (mesoendoderm induction, patterning of the nervous system, and determination of dorsal- ventral axis in vertebrate embryos).

Control Suggestions

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

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. 2012. Partial promoter substitutions generating transcriptional sentinels of diverse signaling pathways in embryonic stem cells and mice. Dis Model Mech 5(6):956-66PubMed: 22888097 MGI: J:195112

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Genetics

Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn}

Allele Symbol: Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn}

Allele Name	targeted mutation 2.1, Mark A Magnussen
Allele Type	Targeted
Allele Synonym(s)	-228/AR8-TA-mCherry; Rosa26^{(228.AR8.TA.mCherry)Hri}
Gene Symbol and Name	Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano
Gene Synonym(s)
Promoter	mixl1, Mix paired homeobox, frog, western clawed
Expressed Gene	RFP, Red Fluorescent Protein, coral
Site of Expression	Red fluorescent protein, mCherry, is expressed in cells and tissues involved in the Nodal signal transduction pathway (mesoendoderm induction, patterning of the nervous system, and determination of dorsal- ventral axis in vertebrate embryos).
Strain of Origin	129S6/SvEvTac
Chromosome	6
General Note	This allele was generate from Gt(ROSA)26Sor^tm1Mgn.
Molecular Note	This allele was created by RMCE in Gt(ROSA)26Sor^tm1Mgn. The original selections cassettes (puromycin N-acetyltransferase and a truncated form of HSV1 thymidine kinase and EM7-driven neomycin resistance) were replaced by a cassette composed of ROSA26 upstream sequence deleted from the start codon to -228, 8 copies of the 50-bp activin response element (ARE) from the Xenopus laevis Mix1b promoter controlling expression of the mCherry reporter. An upstream FLP flanked hygromycin selection cassette was subsequently removed by flp recombinase.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Research Tools
- Developmental Biology Research
- Fluorescent Proteins

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn}/Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn}
involves: 129S6/SvEvBrd * C57BL/6J * SJL

mammalian phenotype

no phenotypic analysis
- Normal - expression studies only

References

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. 2012. Partial promoter substitutions generating transcriptional sentinels of diverse signaling pathways in embryonic stem cells and mice. Dis Model Mech 5(6):956-66PubMed: 22888097 MGI: J:195112

Additional - Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn} related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Gt(ROSA)26Sor
- Genotyping resources and troubleshooting
Breeding Considerations

While maintaining a live colony, these mice are bred as heterozygotes. The Donating Investigator has not attempted to make this strain homozygous for the targeted allele.
- Additional Breeding and Husbandry Support
Citation
When using the B6.129S6(SJL)-Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn}/Mmjax mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #36911 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

The Jackson Laboratory's Genotype Promise

The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.

Terms Of Use

General Terms and Conditions

See MMRRC for Additional Conditions of Distribution

Questions about Terms of Use

Licensing Information

Phone: 207-288-6470

Email: TechTran@jax.org

Donating Investigator

Genetic overview

Research Applications

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Gt(ROSA)26Sortm2.1(mix1b-mCherry)Mgn

Allele Symbol: Gt(ROSA)26Sortm2.1(mix1b-mCherry)Mgn

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

Genotype: Gt(ROSA)26Sortm2.1(mix1b-mCherry)Mgn/Gt(ROSA)26Sortm2.1(mix1b-mCherry)Mgninvolves: 129S6/SvEvBrd * C57BL/6J * SJL

mammalian phenotype

References

Additional - Gt(ROSA)26Sortm2.1(mix1b-mCherry)Mgn related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

The Jackson Laboratory's Genotype Promise

Terms Of Use

Licensing Information

Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn}

Allele Symbol: Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn}

Genotype: Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn}/Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn}
involves: 129S6/SvEvBrd * C57BL/6J * SJL

Additional - Gt(ROSA)26Sor^{tm2.1(mix1b-mCherry)Mgn} related