In these double mutant mice, MEF2C (myocyte enhancer factor 2C) is biotinylated, allowing for specific imunoprecipitation of both MEF2C and the proteins MEF2C interacts with.
William T Pu, Children's Hospital Boston, Harvard MS
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Not Applicable) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Allele Type | Gene Symbol | Gene Name |
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Targeted (Inserted expressed sequence) | Mef2c | myocyte enhancer factor 2C |
These biallelic mutant mice possess a FLAG/bio epitope tag downstream of the 3' UTR of Mef2c.
MEF2C (myocyte enhancer factor 2C) is a member of the MADS box family of transcription enhancer factors. MEF2C has both trans-activating and DNA binding properties and functions in myogenesis, development of the anterior heart field, neural crest and craniofacial development, and neurogenesis.
These mice also contain the Gt(ROSA)26Sortm1.1(birA)Mejr allele, which expresses HA (hemagglutinin) tagged bacterial birA, bifunctional protein (biotin ligase). In the homozygous state, Mef2ctm1.1Wtp is perinatal lethal.
In these double mutant mice, MEF2C 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 Mef2c 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 Mef2c-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 carrying 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 |
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Site of Expression | |
Expressed Gene | FLAG, FLAG octapeptide, |
Site of Expression |
Allele Name | targeted mutation 1.1, Dies Meijer |
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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 |
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Allele Type | Targeted (Inserted expressed sequence) |
Allele Synonym(s) | CVDC/B2B program mutation; Mef2c-flbio |
Gene Symbol and Name | Mef2c, myocyte enhancer factor 2C |
Gene Synonym(s) | |
Expressed Gene | FLAG, FLAG octapeptide, |
Strain of Origin | 129S4/SvJae |
Chromosome | 13 |
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 Mef2c 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 heterozygous for the Mef2c allele and homozygous for the ROSA allele. In the homozygous state, Mef2ctm1.1Wtp is perinatal lethal.
When using the Rosa26BirA; Mef2c-flbio mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #37512 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.
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