Overview

Also Known As:Rosa26^BirA; Srf-flbio

In these double mutant mice, SRF (serum response factor) is biotinylated, allowing for specific imunoprecipitation of both SRF and the proteins SRF interacts with.

Donating Investigator

William T Pu, Children's Hospital Boston, Harvard MS

Genetic overview

Genetic Background	Generation

Gt(ROSA)26Sor^{tm1.1(birA)Mejr}

Allele Type	Gene Symbol	Gene Name
Targeted (Not Applicable)	Gt(ROSA)26Sor	gene trap ROSA 26, Philippe Soriano

Srf^tm1.1Wtp

Allele Type	Gene Symbol	Gene Name
Targeted (Inserted expressed sequence)	Srf	serum response factor

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Research Applications

Research Tools

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Details

Detailed Description

These biallelic mutant mice possess a FLAG/bio epitope tag downstream of the 3' UTR of Srf.

SRF (serum response factor) is a member of MADS box superfamily of transcription factors. It binds to the serum response element (SRE) in the promoter region of target genes and regulates the activity of many immediate-early genes. SRF has a role in cell cycle regulation, apoptosis, cell growth and cell differentiation.

These mice also contain the Gt(ROSA)26Sor^{tm1.1(birA)Mejr} allele, which expresses HA (hemagglutinin) tagged bacterial birA, bifunctional protein (biotin ligase). Mice homozygous for both alleles are viable and fertile.

In these double mutant mice, SRF is biotinylated, allowing for its specific imunoprecipitation as well as any bound proteins.

Development

A targeting vector was designed to insert a FLAG/bio epitope tag and a frt-flanked neomycin resistance (neo) cassette at the endogenous Srf stop codon and upstream of the 3' UTR.

The construct was electroporated into 129S4/SvJae-derived J1 embryonic stem (ES) cells and correctly targeted ES cells were injected into blastocysts. The resulting chimeric mice were bred to outbred Swiss Webster mice. Offspring were bred with Tg(ACTFLPe)9205Dym transgenic mice to delete the neo cassette. The resulting Srf-flbio mice were maintained on a Swiss Webster background.

The Gt(ROSA)26Sor^{tm1.1(birA)Mejr} allele was created using an HA (hemagglutinin) tagged bacterial birA, bifunctional protein (biotin ligase), gene inserted into exon 3 of the rabbit beta-globin gene. The entire birA ORF, with the rabbit beta globin splice acceptor and transcription termination sequences were excised to generate a targeting vector that also contained a floxed PGK- puromycin selection cassette. This targeting vector was inserted into the first intron of the Gt(ROSA)26Sor gene. The construct was electroporated into 129P2/OlaHsd derived IB10 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to FVB/N mice. The mice were intercrossed for 3 generations before being crossed to a Cre recombinase expressing strain (on an unknown genetic background) to remove the loxP-flanked PGK-puromycin selection cassette. A single loxP site remains. The Cre transgene was removed by breeding.

Mice homozygous for both alleles mice were maintained by the Cardiovascular Development Consortium (CvDC), an NIH-funded consortium within the Bench-to-Bassinett (B2B) program.

Upon arrival, mice were bred to FVB/NJ inbred mice for at least one generation to establish the colony.

Expression Data

Expressed Gene	birA, biotin ligase), bacteria
Site of Expression
Expressed Gene	Srf, serum response factor, mouse, laboratory
Expressed Gene	FLAG, FLAG octapeptide,
Site of Expression

Control Suggestions

None Available

Additional Information

Considerations for Choosing Controls

Genetics

Gt(ROSA)26Sor^{tm1.1(birA)Mejr}

Allele Symbol: Gt(ROSA)26Sor^{tm1.1(birA)Mejr}

Allele Name	targeted mutation 1.1, Dies Meijer
Allele Type	Targeted (Not Applicable)
Allele Synonym(s)	bira; Rosa26^BirA
Gene Symbol and Name	Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano
Gene Synonym(s)
Expressed Gene	birA, biotin ligase), bacteria
Strain of Origin	129P2/OlaHsd
Chromosome	6
Molecular Note	An HA (hemagglutinin) tagged E. coli bacterial birA, bifunctional protein (biotin ligase), gene was inserted into exon 3 of the rabbit beta-globin gene. The entire birA ORF, with the rabbit beta globin splice acceptor and transcription termination sequences were excised to generate a targeting vector that also contained a floxed PGK- puromycin selection cassette. This targeting vector was inserted into the first intron of the Gt(ROSA)26Sor gene. Cre-mediated recombination removed the floxed puromycin cassette. Western blot of mutants demonstrated the expression of birA in all tissues tested, though levels of expression varied from tissue to tissue.
Mutations Made By	Dies Meijer, University of Edinburgh Medical School

Srf^tm1.1Wtp

Allele Symbol: Srf^tm1.1Wtp

Allele Name	targeted mutation 1.1, William T Pu
Allele Type	Targeted (Inserted expressed sequence)
Allele Synonym(s)	CVDC/B2B program mutation; Srf-flbio
Gene Symbol and Name	Srf, serum response factor
Gene Synonym(s)
Expressed Gene	Srf, serum response factor, mouse, laboratory
Expressed Gene	FLAG, FLAG octapeptide,
Strain of Origin	129S4/SvJae
Chromosome	17
Molecular Note	A targeting vector was designed to insert a FLAG/bio epitope tag and a frt-flanked neomycin resistance (neo) cassette at the endogenous Srf stop codon and upstream of the 3' UTR. FLP mediated recombination removed the FRT-flanked neo cassette.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Research Tools
- Genetics Research
  - Tissue/Cell Markers

Mammalian Phenotype Terms by Genotype

References

Additional - Gt(ROSA)26Sor^{tm1.1(birA)Mejr} related

Additional - Srf^tm1.1Wtp related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Separated MCA:Gt(ROSA)26Sortm1(birA)Mejr MCA
- Standard PCR:Srf
- Genotyping resources and troubleshooting
Breeding Considerations

While maintaining a live colony, these mice are bred as homozygotes.
- Additional Breeding and Husbandry Support
Citation
When using the Rosa26^BirA; Srf-flbio mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #37511 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

Also Known As:Rosa26BirA; Srf-flbio

Donating Investigator

Genetic overview

Research Applications

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Gt(ROSA)26Sortm1.1(birA)Mejr

Allele Symbol: Gt(ROSA)26Sortm1.1(birA)Mejr

Srftm1.1Wtp

Allele Symbol: Srftm1.1Wtp

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

References

Additional - Gt(ROSA)26Sortm1.1(birA)Mejr related

Additional - Srftm1.1Wtp 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

Also Known As:Rosa26^BirA; Srf-flbio

Gt(ROSA)26Sor^{tm1.1(birA)Mejr}

Allele Symbol: Gt(ROSA)26Sor^{tm1.1(birA)Mejr}

Srf^tm1.1Wtp

Allele Symbol: Srf^tm1.1Wtp

Additional - Gt(ROSA)26Sor^{tm1.1(birA)Mejr} related

Additional - Srf^tm1.1Wtp related