BRafCA is a knock-in allele of human BRaf which expresses normal BRaf prior to Cre recombinase exposure. Cre-mediated recombination results in the BRafVE allele, which expresses the constitutively active BRAFV600E mutant protein. In conjunction with Cre-expressing mouse lines, these BRafCA mice may be useful in studies of human malignancies, as well as for studying the RAS-activated RAF/MEK/ERK mitogen-activated protein kinase signaling pathway and pleiotropic regulators of the aberrant cancer cell physiology.
IMR Colony, The Jackson Laboratory
Genetic Background | Generation |
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?+F12pN1F13
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), No functional change) | Braf | Braf transforming gene |
Mice homozygous for the Cre-activated BRaf mutant allele (BRafCA) are viable and fertile. Mice express the transcript encoded by endogenous exons 1-14 and loxP-flanked human exons 15-18 (BRafCA) prior to Cre recombinase exposure. Homozygous mice have "normal" BRaf expression, and no abnormalities are reported for homozygous BRafCA mice.
Following Cre-mediated excision of the floxed sequences (human BRAF exons 15-18 and polyA signal), the transcripts are subsequently generated using the downstream mutant exon 15 (modified with a V600E amino acid substitution associated with constitutively active BRAFV600E in human cancers) and the endogenous downstream exons 16-18. The resulting BRafVE transcripts are subject to normal patterns of alternate splicing and differential exon usage, and BRafVE expression is observed at physiological levels. To discriminate the expression of the various BRaf mRNAs in heterozygous mice, silent restriction enzyme polymorphisms were incorporated into the exon 15 sequences of BRafCA (BamHI) and BRafVE (XbaI).
For example, when crossed to a strain expressing tamoxifen-inducible Cre recombinase (see Stock No. 017585), this mutant mouse strain may be useful in studies of lung tumors.
The BRafCA mutant mice were created by Dr. Martin McMahon (University of California, San Francisco). The BRafCA targeted mutation was designed to replace the endogenous Braf exon 15 with (from 5' to 3') a loxP site, the cDNA sequence encoded by human BRAF exons 15-18 (with a silent BamHI restriction site polymorphism in exon 15), the mouse Braf polyA sequences, a PGK-neo cassette (with triple transcriptional stop and SV40 polyA signal), a second loxP site, and a mouse exon 15 sequence modified via site-directed mutagenesis to harbor both a V600E amino acid substitution (associated with the constitutively active BRAFV600E mutation in human cancers) and also a silent XbaI restriction site polymorphism. The targeting vector was microinjected into 129Sv/OLA embryonic stem (ES) cells, and correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric mice were bred with FVB/N mice to establish the colony. BRafCA mutant mice were subsequently maintained on a C57BL/6 congenic background.
This BRafCA strain was established at The Jackson Laboratory in 2012 from the same project that assembled the BRafCA PtenloxP Tyr::CreERT2 triple mutant line (Stock No. 013590). Mice identified by genotype with the BRafCA allele, without the PtenloxP allele and without the Tyr::CreERT2 transgene (confirmed September 2015) were used as founders for this BRafCA strain (Stock No. 017837).
Expressed Gene | BRAF, B-Raf proto-oncogene, serine/threonine kinase, human |
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Site of Expression | intranasal instillation of an adenovirus expressing Cre recombinase results in benign lung tumors that only rarely progress to adenocarcinoma. |
Allele Name | targeted mutation 1, Martin McMahon |
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Allele Type | Targeted (Conditional ready (e.g. floxed), No functional change) |
Allele Synonym(s) | BrafCA; BRAFF-V600E; BRAFV600Eca |
Gene Symbol and Name | Braf, Braf transforming gene |
Gene Synonym(s) | |
Expressed Gene | BRAF, B-Raf proto-oncogene, serine/threonine kinase, human |
Site of Expression | intranasal instillation of an adenovirus expressing Cre recombinase results in benign lung tumors that only rarely progress to adenocarcinoma. |
Strain of Origin | 129P2/OlaHsd |
Chromosome | 6 |
Molecular Note | A targeting vector was designed to insert a loxP site and the human BRAF cDNA covering exons 15-18 into intron 14, along with a floxed neo cassette. A modified exon 15 was engineered into the vector to encode Braf |
Mutations Made By | Martin McMahon, University of California, San Francisco |
When maintaining a live colony, these mice can be bred as homozygous for the Braftm1Mmcm allele.
When using the BRafCA mouse strain in a publication, please cite the originating article(s) and include JAX stock #017837 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Heterozygous for Braf<tm1Mmcm> |
Frozen Mouse Embryo | B6.129P2(Cg)-Braf<tm1Mmcm>/J | $2595.00 |
Frozen Mouse Embryo | B6.129P2(Cg)-Braf<tm1Mmcm>/J | $2595.00 |
Frozen Mouse Embryo | B6.129P2(Cg)-Braf<tm1Mmcm>/J | $3373.50 |
Frozen Mouse Embryo | B6.129P2(Cg)-Braf<tm1Mmcm>/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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