In these double mutant mice, SRF (serum response factor) is biotinylated, allowing for specific imunoprecipitation of both SRF and the proteins SRF interacts with.
William T Pu, Children's Hospital Boston, Harvard MS
Genetic Background | Generation |
---|---|
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Not Applicable) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Inserted expressed sequence) | Srf | serum response factor |
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)26Sortm1.1(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.
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)26Sortm1.1(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.
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 |
Allele Name | targeted mutation 1.1, Dies Meijer |
---|---|
Allele Type | Targeted (Not Applicable) |
Allele Synonym(s) | bira; Rosa26BirA |
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 |
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. |
While maintaining a live colony, these mice are bred as homozygotes.
When using the Rosa26BirA; Srf-flbio mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #37511 in your Materials and Methods section.
Facility Barrier Level Descriptions
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.
What information were you hoping to find through your search?
How easy was it to find what you were looking for?
We may wish to follow up with you. Enter your email if you are happy for us to connect and reachout to you with more questions.
Please Enter a Valid Email Address
Thank you for sharing your feedback! We are working on improving the JAX Mice search. Come back soon for exciting changes.