The R26StopFLMEK1DD conditional allele is targeted to the Gt(ROSA)26Sor locus and has a loxP-flanked Neo-STOP cassette preventing transcription of the downstream bicistronic sequences (MEK1DD [a mutant form of rat MAPKK1 rendered constitutively active by two serine->aspartic acid substitutions (S218D/S222D) within the catalytic domain] and EGFP). These R26StopFLMEK1DD mice allow inducible constitutive activity of MAP Kinase signal transduction pathways that mediate various cellular activities, including gene expression, mitosis, differentiation, proliferation, cell survival, and cell cycle progression.
Klaus Rajewsky, Max Delbruck Centre for Molecular Medicine
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), Reporter, Constitutively active, Inserted expressed sequence) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Mice homozygous for the R26StopFLMEK1DD conditional allele are viable and fertile, with a loxP-flanked Neo-STOP cassette preventing transcription of the downstream bicistronic sequences (MEK1DD [a mutant form of rat MAPKK1 rendered constitutively active by two serine->aspartic acid substitutions (S218D/S222D) within the catalytic domain] and EGFP). When bred to mice that express Cre recombinase, offspring will have the STOP cassette deleted and subsequent expression of the MEK1DD signal molecule and EGFP fluorescence in the cre-expressing cells. Expression of MEK1DD leads to constitutive activity of MAP Kinase signal transduction pathways that mediate various cellular activities, including gene expression, mitosis, differentiation, proliferation, cell survival, and cell cycle progression. Of note, breeding these mice to an FLP-expressing strain will result in removal of the frt-flanked IRES-EGFP cassette.
A targeting vector designed with (from 5' to 3') a splice acceptor site, a loxP-flanked Neo-STOP cassette, a cDNA sequence encoding rat MEK1DD (a mutant form of MAPKK1 rendered constitutively active by two serine->aspartic acid substitutions (S218D/S222D) within the catalytic domain], an frt-flanked internal ribosomal entry site-enhanced green fluorescent protein cassette (IRES-EGFP), and bovine polyadenylation signal. This transgene construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus via electroporation into C57BL/6-derived embryonic stem (ES) cells. Correctly targeted ES cells were selected and chimeric animals were bred to C57BL/6 to generate the mutant colony. The donating investigator reports that these R26StopFLMEK1DD mice were maintained on a C57BL/6 genetic background (see SNP note below) as a homozygous colony for many generations prior to arrival at The Jackson Laboratory Repository. Upon arrival, mutant mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. In some of the mice, one marker on chromosome 11 (~8.4 Mbp) and one marker on chromosome 17 (~34.3 Mbp) were not fixed to the C57BL/6 genetic background. The source of this is not known. In addition, at least 3 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating in all the rederived mice. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.
Expressed Gene | Map2k1, mitogen activated protein kinase kinase 1, rat |
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Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | Cre excision of the stop signal results in expression of EGFP and a constitutively active form of rat MAPKK1 (MEK1DD) in cre-expressing tissues. |
Allele Name | targeted mutation 8, Klaus Rajewsky |
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Allele Type | Targeted (Conditional ready (e.g. floxed), Reporter, Constitutively active, Inserted expressed sequence) |
Allele Synonym(s) | MEk1DD; R26Stop flox MEK1DD; R26StopFLMEK1DD; R26Stop-flox-MEK1DD; ROSAMEK1* |
Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
Gene Synonym(s) | |
Expressed Gene | Map2k1, mitogen activated protein kinase kinase 1, rat |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | Cre excision of the stop signal results in expression of EGFP and a constitutively active form of rat MAPKK1 (MEK1DD) in cre-expressing tissues. |
Strain of Origin | C57BL/6 |
Chromosome | 6 |
Molecular Note | A targeting vector designed with (from 5' to 3') a splice acceptor site, a loxP-flanked Neo-STOP cassette, a cDNA sequence encoding rat MEK1DD (a mutant form of MAPKK1 rendered constitutively active by two serine->aspartic acid substitutions (S218D/S222D) within the catalytic domain], an frt-flanked internal ribosomal entry site-enhanced green fluorescent protein cassette (IRES-EGFP), and bovine polyadenylation signal. This transgene construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. |
Mutations Made By | Klaus Rajewsky, Max Delbruck Centre for Molecular Medicine |
When maintaining a live colony, homozygous mice may be bred together.
When using the C57BL/6-Gt(ROSA)26Sortm8(Map2k1*,EGFP)Rsky/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #012352 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous or wildtype for Gt(ROSA)26Sor<tm8(Map2k1*,EGFP)Rsky> |
Frozen Mouse Embryo | C57BL/6-Gt(ROSA)26Sor<tm8(Map2k1*EGFP)Rsky>/J | $2595.00 |
Frozen Mouse Embryo | C57BL/6-Gt(ROSA)26Sor<tm8(Map2k1*EGFP)Rsky>/J | $2595.00 |
Frozen Mouse Embryo | C57BL/6-Gt(ROSA)26Sor<tm8(Map2k1*EGFP)Rsky>/J | $3373.50 |
Frozen Mouse Embryo | C57BL/6-Gt(ROSA)26Sor<tm8(Map2k1*EGFP)Rsky>/J | $3373.50 |
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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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