Overview

Also Known As: BRaf^CA, BRAF^F-V600E

BRaf^CA is a knock-in allele of human BRaf which expresses normal BRaf prior to Cre recombinase exposure. Cre-mediated recombination results in the BRaf^VE allele, which expresses the constitutively active BRAF^V600E mutant protein. In conjunction with Cre-expressing mouse lines, these BRaf^CA 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.

Donating Investigator

IMR Colony, The Jackson Laboratory

Genetic overview

Genetic Background	Generation
	?+F12pN1F7 (2018-04-20 00:00:00)

Braf^tm1Mmcm

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), No functional change)	Braf	Braf transforming gene

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Research Applications

Cell Biology Research
Cancer Research
Research Tools
Immunology, Inflammation and Autoimmunity Research

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Base Price

Starting at:

$278.00 Domestic price for female 4-week

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Details

Detailed Description

Mice homozygous for the Cre-activated BRaf mutant allele (BRaf^CA) are viable and fertile. Mice express the transcript encoded by endogenous exons 1-14 and loxP-flanked human exons 15-18 (BRaf^CA) prior to Cre recombinase exposure. Homozygous mice have "normal" BRaf expression, and no abnormalities are reported for homozygous BRaf^CA 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 BRAF^V600E in human cancers) and the endogenous downstream exons 16-18. The resulting BRaf^VE transcripts are subject to normal patterns of alternate splicing and differential exon usage, and BRaf^VE 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 BRaf^CA (BamHI) and BRaf^VE (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.

Development

The BRaf^CA mutant mice were created by Dr. Martin McMahon (University of California, San Francisco). The BRaf^CA 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 BRAF^V600E 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. BRaf^CA mutant mice were subsequently maintained on a C57BL/6 congenic background.

This BRaf^CA strain was established at The Jackson Laboratory in 2012 from the same project that assembled the BRaf^CA Pten^loxP Tyr::CreER^T2 triple mutant line (Stock No. 013590). Mice identified by genotype with the BRaf^CA allele, without the Pten^loxP allele and without the Tyr::CreER^T2 transgene (confirmed September 2015) were used as founders for this BRaf^CA strain (Stock No. 017837).

Expression Data

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.

Control Suggestions

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Dankort D; Filenova E; Collado M; Serrano M; Jones K; McMahon M. 2007. A new mouse model to explore the initiation, progression, and therapy of BRAFV600E-induced lung tumors. Genes Dev 21(4):379-84PubMed: 17299132 MGI: J:121715

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Genetics

Braf^tm1Mmcm

Allele Symbol: Braf^tm1Mmcm

Allele Name	targeted mutation 1, Martin McMahon
Allele Type	Targeted (Conditional ready (e.g. floxed), No functional change)
Allele Synonym(s)	targeted mutation 1, Martin McMahon; Braf^tm1Mmcm
Gene Symbol and Name	Braf, Braf transforming gene
Gene Synonym(s)	B-RAF1; 9930012E13Rik; expressed sequence AA387315; NS7; expressed sequence AA120551; D6Ertd631e; Braf-2; BRAF1; Braf2; RAFB1; RIKEN cDNA 9930012E13 gene; 9930012E13Rik; AA473386; expressed sequence AA473386; DNA segment, Chr 6, ERATO Doi 631, expressed; Braf transforming gene 2; AA387315; AA120551; expressed sequence C87398; Braf-2; Braf2; C87398; D6Ertd631e; B-raf
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 and a silent XbaI restriction site. RT-PCR confirmed that mutant transcript was expressed at the same level as wild-type, and that the V600E mutation was not detected.
Mutations Made By	Martin McMahon, University of California, San Francisco

Disease/Phenotype

Disease Terms

Models with phenotypic similarity to human diseases where etiology is unknown or involving genes where ortholog is unknown.

Langerhans-cell histiocytosis

Research Areas By Genotype

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

Cell Biology Research
- Signal Transduction
- Transcriptional Regulation
Cancer Research
- Oncogenes
  - control strain
- Other
  - tumor metastasis
Research Tools
- Developmental Biology Research
  - Cre-lox System
- Genetics Research
  - Tissue/Cell Markers: Cre-lox System
  - Mutagenesis and Transgenesis: Cre-lox System
- Cancer Research
- Cre-lox System
  - loxP-flanked Sequences
Immunology, Inflammation and Autoimmunity Research
- Intracellular Signaling Molecules

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Braf^tm1Mmcm/Braf⁺
involves: 129P2/OlaHsd

mortality/aging

premature death
- 7-8 weeks after adenoviral Cre intranasal administration to 6- to 8-week old mutants, treated animals are euthanized because they show labored breathing and general wasting

normal phenotype

no abnormal phenotype detected
- Normal - mice are fertile and born at Mendelian ratios, but no further phenotypic analysis is provided

respiratory system phenotype

increased lung adenocarcinoma incidence
- only 2 mice (of 57) that develop adenomas show evidence of progression to adenocarcinoma; cells from these mice display nuclear enlargement, hyperchromasia, and contour irregularities

increased lung adenoma incidence
- 7 weeks after high dose Cre treatment, evidence of multiple adenomatous tumor formation is present in mouse lungs, while no tumors are detected in untreated mice after 6 months
- after lower dose Cre treatement, adenomas do not grow beyond ~900 um in diameter
- at ~14 weeks, papillary adenomas undergo changes at periphery; cells change such that they have more cytoplasm and become eosinophilic
- lesions arise in alveolar ducts expanding outward and around broncioles
- papillary adenomas are detected 6-8 weeks after lower dose adenoviral Cre treatment; lesions seem to increase in size and number

increased lung tumor incidence
- 4 weeks after Cre treatment, mice are treated for 28 days with a pharmacological inhibitor of MEK1/2 which prevents almost all tumor development, while vehicle-treated animals show numerous lung tumors
- cellular origins of cancerous are ambiguous aslesions are Clara Cell antigen-negative, but majority of cells express Surfactant protein C as early as 2 weeks after Cre treatment (CCA and SP-C are both markers of alveolar type II pneumocytes)

lung epithelium hyperplasia
- 4 weeks after Cre administration, lung epithelium hyperplasia is observed
- at earlier times after Cre treatment, hyperplastic epithelium shows papillary outgrowths, but these were not seen at earlier time points (2-4 weeks post-treatment)

neoplasm

increased lung adenocarcinoma incidence
- only 2 mice (of 57) that develop adenomas show evidence of progression to adenocarcinoma; cells from these mice display nuclear enlargement, hyperchromasia, and contour irregularities

increased lung adenoma incidence
- 7 weeks after high dose Cre treatment, evidence of multiple adenomatous tumor formation is present in mouse lungs, while no tumors are detected in untreated mice after 6 months
- after lower dose Cre treatement, adenomas do not grow beyond ~900 um in diameter
- at ~14 weeks, papillary adenomas undergo changes at periphery; cells change such that they have more cytoplasm and become eosinophilic
- lesions arise in alveolar ducts expanding outward and around broncioles
- papillary adenomas are detected 6-8 weeks after lower dose adenoviral Cre treatment; lesions seem to increase in size and number

increased lung tumor incidence
- 4 weeks after Cre treatment, mice are treated for 28 days with a pharmacological inhibitor of MEK1/2 which prevents almost all tumor development, while vehicle-treated animals show numerous lung tumors
- cellular origins of cancerous are ambiguous aslesions are Clara Cell antigen-negative, but majority of cells express Surfactant protein C as early as 2 weeks after Cre treatment (CCA and SP-C are both markers of alveolar type II pneumocytes)

Genotype: Braf^tm1Mmcm/Braf⁺ Polr2a^{tm1(cre/ERT2)Bbd}/Polr2a⁺
involves: 129P2/OlaHsd * C57BL/6 * SJL

neoplasm

increased lung adenoma incidence
- with induction of Cre expression from the transgene, multiple adenomatous tumors form in mouse lungs

respiratory system phenotype

increased lung adenoma incidence
- with induction of Cre expression from the transgene, multiple adenomatous tumors form in mouse lungs

Genotype: Braf^tm1Mmcm/Braf⁺ Tg(TPO-cre/ERT2)1139Tyj/0
involves: 129P2/OlaHsd * 129S4/SvJae * C57BL/6 * C57BL/6NTac

endocrine/exocrine gland phenotype

increased thyroid carcinoma incidence
- even with very long latency, extrathyroidal invasion is not seen and cervical lymph node metastases are not seen
- mice administered tamoxifen develop papillary thyroid carcinomas
- tumors show both papillary growth morphology and nuclear features of papillary thyroid carcinoma

neoplasm

increased thyroid carcinoma incidence
- even with very long latency, extrathyroidal invasion is not seen and cervical lymph node metastases are not seen
- mice administered tamoxifen develop papillary thyroid carcinomas
- tumors show both papillary growth morphology and nuclear features of papillary thyroid carcinoma

respiratory system phenotype

respiratory failure
- tamoxifen treated mice generally succumb to respiratory compromise due to tracheal compression by large noninvasive tumors

Genotype: Braf^tm1Mmcm/Braf⁺
involves: 129P2/OlaHsd * 129S4/SvJae * C57BL/6

endocrine/exocrine gland phenotype

increased thyroid carcinoma incidence
- mice with surgical delivery of a cre-expressing adenovirus to the adult thyroid gland develop papillary thyroid carcinoma

neoplasm

increased thyroid carcinoma incidence
- mice with surgical delivery of a cre-expressing adenovirus to the adult thyroid gland develop papillary thyroid carcinoma

Genotype: Braf^tm1Mmcm/Braf⁺ Tg(Tyr-cre/ERT2)13Bos/0
involves: 129P2/OlaHsd * FVB

integument phenotype

skin lesions
- mice treated with 4-hydroxytamoxifen (4-HT) either topically or systemically develop highly pigmented lesions that are detected by 21-28 days after 4-HT administration; these are small papular pigmented lesions that are not malignant
- topical administration of high concentrations of 4-HT occasionally results in periocular lesions on the face and on the glabrous skin of the paws

pigmentation phenotype

abnormal melanocyte morphology
- other melanocytic proliferations form in the dermis with no junctional component in 4-HT treated mice
- pigmented lesions in 4-HT treated mice are frequently located at the dermal-epidermal junction with extension into the dermis, indicating melanocytic nevi

Genotype: Braf^tm1Mmcm/Braf^tm1Mmcm Tg(Tyr-cre/ERT2)13Bos/0
involves: 129P2/OlaHsd * FVB

pigmentation phenotype

abnormal melanocyte morphology
- topical administration of 4-hydroxytamoxifen (4-HT) to the ear, tail or flank results in the development of benign melanocytic hyperplasias that are larger and more highly pigmented than in conditional heterozygotes

Genotype: Braf^tm1Mmcm/Braf^tm1Mmcm Tg(Itgax-cre)1-1Reiz/0
involves: 129P2/OlaHsd * C57BL/6 * CBA

hematopoietic system phenotype

anemia
- severe

enlarged spleen
- severe hepatosplenomegaly

increased dendritic cell number
- expansion of blood circulating dendritic cell precursors and blood circulating dendritic cells
- lesions are associated with massive increase of MHC II+CD11c+ dendritic cells

liver/biliary system phenotype

enlarged liver
- severe hepatosplenomegaly

mortality/aging

premature death
- decrease in lifespan

homeostasis/metabolism phenotype

abnormal chemokine level
- granuloma lesions are associated with an increase in several chemokines and cytokines (Ccl2, Ccl15, Il6, Il10, IFN-gamma, and TGF-beta) indicative of a local cytokine storm

abnormal cytokine level
- granuloma lesions are associated with an increase in several chemokines and cytokines (Ccl2, Ccl15, Il6, Il10, IFN-gamma, and TGF-beta) indicative of a local cytokine storm

immune system phenotype

abnormal chemokine level
- granuloma lesions are associated with an increase in several chemokines and cytokines (Ccl2, Ccl15, Il6, Il10, IFN-gamma, and TGF-beta) indicative of a local cytokine storm

abnormal cytokine level
- granuloma lesions are associated with an increase in several chemokines and cytokines (Ccl2, Ccl15, Il6, Il10, IFN-gamma, and TGF-beta) indicative of a local cytokine storm

enlarged lymph nodes

enlarged spleen
- severe hepatosplenomegaly

granulomatous inflammation
- granuloma lesions contain massive CD11c+ langerin+ infiltrates, a large number of macrophages, NK cells, B cells, and T cells, with a specific accumulation of regulatory T cells
- histiocytic infiltrates in the skin, liver, spleen, and lungs by 8 weeks of age, resulting in a broad destruction of tissue architecture by 16 weeks of age
- histiocytic lesions exhibit classical granulomatous organization, including multinucleated giant cell formation
- local fibrotic response is seen within the granulomas and surrounding stroma

increased dendritic cell number
- expansion of blood circulating dendritic cell precursors and blood circulating dendritic cells
- lesions are associated with massive increase of MHC II+CD11c+ dendritic cells

increased inflammatory response
- mice rapidly develop an aggressive Langerhans-cells histiocytosis-like disease with 100% penetrance

References

Dankort D; Filenova E; Collado M; Serrano M; Jones K; McMahon M. 2007. A new mouse model to explore the initiation, progression, and therapy of BRAFV600E-induced lung tumors. Genes Dev 21(4):379-84PubMed: 17299132 MGI: J:121715

Additional - Braf^tm1Mmcm related

Technical Support

Contact Technical Support

Genotyping Protocols
- High Resolution Melting: Braf^tm1Mmcm
- Standard PCR: Braf^tm1Mmcm
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, these mice can be bred as homozygous for the Braf^tm1Mmcm allele.
- Additional Breeding and Husbandry Support
Mating System
- Homozygote x Homozygote
Citation
When using the BRaf^CA mouse strain in a publication, please cite the originating article(s) and include JAX stock #017837 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

AX18 (Maximum)

Pricing & Availability

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Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service	Genotype	Price
	Heterozygous for Braf<tm1Mmcm>

We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.

Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation.

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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.

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Also Known As: BRafCA, BRAFF-V600E

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Braftm1Mmcm

Allele Symbol: Braftm1Mmcm

Disease Terms

Models with phenotypic similarity to human diseases where etiology is unknown or involving genes where ortholog is unknown.

Research Areas By Genotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Braftm1Mmcm/Braf+involves: 129P2/OlaHsd

mortality/aging

normal phenotype

respiratory system phenotype

neoplasm

Genotype: Braftm1Mmcm/Braf+ Polr2atm1(cre/ERT2)Bbd/Polr2a+involves: 129P2/OlaHsd * C57BL/6 * SJL

neoplasm

respiratory system phenotype

Genotype: Braftm1Mmcm/Braf+ Tg(TPO-cre/ERT2)1139Tyj/0involves: 129P2/OlaHsd * 129S4/SvJae * C57BL/6 * C57BL/6NTac

endocrine/exocrine gland phenotype

neoplasm

respiratory system phenotype

Genotype: Braftm1Mmcm/Braf+involves: 129P2/OlaHsd * 129S4/SvJae * C57BL/6

endocrine/exocrine gland phenotype

neoplasm

Genotype: Braftm1Mmcm/Braf+ Tg(Tyr-cre/ERT2)13Bos/0involves: 129P2/OlaHsd * FVB

integument phenotype

pigmentation phenotype

Genotype: Braftm1Mmcm/Braftm1Mmcm Tg(Tyr-cre/ERT2)13Bos/0involves: 129P2/OlaHsd * FVB

pigmentation phenotype

Genotype: Braftm1Mmcm/Braftm1Mmcm Tg(Itgax-cre)1-1Reiz/0involves: 129P2/OlaHsd * C57BL/6 * CBA

hematopoietic system phenotype

liver/biliary system phenotype

mortality/aging

homeostasis/metabolism phenotype

immune system phenotype

References

Additional - Braftm1Mmcm related

Genotyping Protocols

Breeding Considerations

Mating System

Citation

Animal Health Reports

Live Mouse

Cryorecovery - Pricing

Related Products and Services

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms of Use

Additional Use Restrictions Apply

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Also Known As: BRaf^CA, BRAF^F-V600E

Braf^tm1Mmcm

Allele Symbol: Braf^tm1Mmcm

Genotype: Braf^tm1Mmcm/Braf⁺
involves: 129P2/OlaHsd

Genotype: Braf^tm1Mmcm/Braf⁺ Polr2a^{tm1(cre/ERT2)Bbd}/Polr2a⁺
involves: 129P2/OlaHsd * C57BL/6 * SJL

Genotype: Braf^tm1Mmcm/Braf⁺ Tg(TPO-cre/ERT2)1139Tyj/0
involves: 129P2/OlaHsd * 129S4/SvJae * C57BL/6 * C57BL/6NTac

Genotype: Braf^tm1Mmcm/Braf⁺
involves: 129P2/OlaHsd * 129S4/SvJae * C57BL/6

Genotype: Braf^tm1Mmcm/Braf⁺ Tg(Tyr-cre/ERT2)13Bos/0
involves: 129P2/OlaHsd * FVB

Genotype: Braf^tm1Mmcm/Braf^tm1Mmcm Tg(Tyr-cre/ERT2)13Bos/0
involves: 129P2/OlaHsd * FVB

Genotype: Braf^tm1Mmcm/Braf^tm1Mmcm Tg(Itgax-cre)1-1Reiz/0
involves: 129P2/OlaHsd * C57BL/6 * CBA

Additional - Braf^tm1Mmcm related