Overview

Also Known As: HEB^flox E2A^flox

HEB^f E2A^f mice allow Cre-recombinase inducible deletion of the two basic helix-loop-helix (bHLH) E-protein family genes HEB and E2A. These mice may be useful for studying B and T lymphocyte development and lymphoid malignancies.

Donating Investigator

Yuan Zhuang, Duke University Medical Center

Genetic overview

Genetic Background	Generation

Tcf12^tm3Zhu

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), No functional change)	Tcf12	transcription factor 12

Tcf3^tm4Zhu

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), No functional change)	Tcf3	transcription factor 3

View Genetics

Research Applications

Research Tools
Immunology, Inflammation and Autoimmunity Research
Virology Research
Internal/Organ Research
Cancer Research
Hematological Research
Developmental Biology Research
Neurobiology Research

View All Research Applications

Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

These HEB^f E2A^f mice harbor two mutations, the HEB floxed allele (Tcf12^tm3Zhu) and E2A floxed allele (Tcf3^tm4Zhu); each with its basic helix-loop-helix domain flanked by loxP sites.

The transcription factors E2A and HEB (members of the E protein family) have been shown to play essential roles in lymphocyte development, and mutations/deficiencies are associated with lymphoid malignancies. Mice homozygous for both floxed alleles are viable and fertile with no reported abnormalities. When bred to mice that express Cre recombinase, the resulting offspring may be useful in generating tissue-specific alterations in B cell and T cell development. The E2A^f allele also expresses β-galactosidase (lacZ) following exposure to Cre recombinase. Specific examples are described below.

HEB^f E2A^f mice may be bred to animals expressing Cre recombinase in different stages of T cell development. When bred to mice expressing Cre recombinase in the very late double-negative stage of T cell development (CD4-Cre transgenic mice; see Stock No. 017336), the resulting mice allow knockout of these two genes before the double-positive stage of T-cell development. When bred to mice expressing Cre recombinase later in T cell development (such as Lck-Cre transgenic mice; Stock No. 012837), the resulting mice allow knockout of these two genes after positive selection in the thymus.

Development

These HEB^f E2A^f mice harbor two mutations; the HEB floxed allele (Tcf12^tm3Zhu) and E2A floxed allele (Tcf3^tm4Zhu).

The HEB^flox (or HEB^f) targeting construct was designed by Dr. Yuan Zhuang (Duke University Medical Center) to insert a loxP site upstream of the basic helix-loop-helix encoding sequence (exon 18), as well as a loxP-flanked PGK-neo cassette downstream of exon 18 of the transcription factor 12 gene (Tcf12) on chromosome 9. The construct was electroporated into undisclosed embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a Cre-recombinase expression vector. ES cells retaining the floxed exon 18 and with the PGK-neo removed were identified and injected into recipient blastocysts. The chimeric animals were bred to undisclosed mice to establish the HEB^f colony. The donating investigator reports that the HEB^f colony was backcrossed with C57BL/6J more than ten generations (see SNP note below). The resulting C57BL/6J-congenic HEB^f mice were then used as described below.

The E2A^loxp (or E2A^f) targeting construct was designed by Dr. Yuan Zhuang (Duke University Medical Center) to insert a loxP site into an intron upstream of the basic helix-loop-helix encoding sequence (common to both E12 and E47 isoforms), as well as a PGK-neo, loxP site, splice acceptor site, IRES-lacZ and transcription stop signal downstream of the coding region of the transcription factor 3 gene (Tcf3) on chromosome 10. The construct was electroporated into 129S4/SvJaeSor-derived AK7 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric animals were bred to undisclosed mice to establish the E2A^f colony. The donating investigator reports that the E2A^f colony was backcrossed with C57BL/6J more than ten generations (see SNP note below). The resulting C57BL/6J-congenic E2A^f mice were then used as described below.

To generate the double floxed line, Dr. Zhuang bred C57BL/6J-congenic HEB^f mice with C57BL/6J-congenic E2A^f mice (see SNP note below). The HEB^f E2A^f mice were sent to The Jackson Laboratory Repository in 2014. Upon arrival, males were used to cryopreserve sperm. To establish our living mouse colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J inbred females (Stock No. 000664).

In 2015, 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 first generation rederived living colony at The Jackson Laboratory Repository. This revealed 4 of 27 markers were not fixed for C57BL/6 allele-type (i.e., still segregating for 129S1 allele-type markers on chromosomes 3 [~37.3 Mbp], 8 [~14.8 Mbp], 11 [~20.9 Mbp] and 12 [~30.7 Mbp]).
In addition, 4 of the 5 markers that determine C57BL/6J from C57BL/6N revealed some mice were segregating for 129S1 allele-type markers (chromosomes 8 [~15.2 Mbp], 11 [~4.4 Mbp], 15 [~57 Mbp] and 19 [~50 Mbp]), and the 5th marker was C57BL/6J.
Collectively, these data suggest the mice sent to The Jackson Laboratory were on a mixed genetic background (~77% C57BL/6).

Control Suggestions

Approximate Controls

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Wojciechowski J; Lai A; Kondo M; Zhuang Y. 2007. E2A and HEB are required to block thymocyte proliferation prior to pre-TCR expression. J Immunol 178(9):5717-26PubMed: 17442955 MGI: J:129570
Lazorchak AS; Wojciechowski J; Dai M; Zhuang Y. 2006. E2A promotes the survival of precursor and mature B lymphocytes. J Immunol 177(4):2495-504PubMed: 16888011 MGI: J:138355
Zhang B; Lin YY; Dai M; Zhuang Y. 2014. Id3 and Id2 act as a dual safety mechanism in regulating the development and population size of innate-like gammadelta T cells. J Immunol 192(3):1055-63PubMed: 24379125 MGI: J:207300
Pan L; Hanrahan J; Li J; Hale LP; Zhuang Y. 2002. An Analysis of T Cell Intrinsic Roles of E2A by Conditional Gene Disruption in the Thymus. J Immunol 168(8):3923-32PubMed: 11937548 MGI: J:75746

View All References

Genetics

Tcf12^tm3Zhu

Allele Symbol: Tcf12^tm3Zhu

Allele Name	targeted mutation 3, Yuan Zhuang
Allele Type	Targeted (Conditional ready (e.g. floxed), No functional change)
Allele Synonym(s)	targeted mutation 3, Yuan Zhuang; Tcf12^tm3Zhu
Gene Symbol and Name	Tcf12, transcription factor 12
Gene Synonym(s)	ALF1; bHLHb20; REB; CRS3; SCBPA; HEB; HEBAlt; HTF-4; HsT17266; HTF4; ME1; TCF-12; SCBP alpha; p64
Strain of Origin	Not Specified
Chromosome	9
Molecular Note	A floxed neo cassette was inserted into intron 18 and an additional loxP site was inserted into intron 17. The neo cassette was removed by cre-mediated recombination using a transiently expressing cre vector leaving a floxed exon 18.

Tcf3^tm4Zhu

Allele Symbol: Tcf3^tm4Zhu

Allele Name	targeted mutation 4, Yuan Zhuang
Allele Type	Targeted (Conditional ready (e.g. floxed), No functional change)
Allele Synonym(s)	targeted mutation 4, Yuan Zhuang; Tcf3^tm4Zhu
Gene Symbol and Name	Tcf3, transcription factor 3
Gene Synonym(s)	E47; ALF2; Tcfe2a; E2A; A1; E12; expressed sequence AW209082; VDIR; TCF-3; Tcfe2a; expressed sequence AA408400; Pan1; Pan2; bHLHb21; ITF1; transcription factor E2a; AW209082; AA408400; AGM8; p75
Strain of Origin	129S4/SvJaeSor
Chromosome	10
Molecular Note	A targeting construct was engineered to contain a 5' loxP site in an intron upstream of the bHLH sequence, common to both E12 and E47 isoforms, and a 3' loxP site downstream of the endogenous locus. Also included in the construct were neo and lacZ genes, respectively positioned upstream and downstream of the 3' loxP site. The construct was designed such that the lacZ gene will be transcribed only upon recombination of the loxP sites.
Mutations Made By	Yuan Zhuang, Duke University Medical Center

Disease/Phenotype

Disease Terms

Research Areas By Genotype

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

Research Tools
- Cancer Research
  - B cell deficiency
  - production of B cells and antibodies
  - genes regulating lymphoma development
  - B and T cell deficiency, xenograft/transplant host
  - Cre-lox System
  - xenograft/transplant host
  - T cell deficiency
  - production of B and T cells, antibodies, and hybridomas
  - production of T cells and hybridoma
  - specific T cell deficiency
- Immunology, Inflammation and Autoimmunity Research
  - B cell deficiency
  - B cell lymphomas
  - production of B cells and antibodies
  - T Cell Receptor Deficiency
  - T cell deficiency
  - T cell deficiency, xenograft/transplant host
  - production of B cells, antibodies T cell lines, and hybridomas
  - production of T cell lines and hybridomas
  - specific T cell deficiency
  - B and T cell deficiency
- Cre-lox System
  - loxP-flanked Sequences
  - loxP-flanked Sequences: Test/Reporter
- lacZ Expression
- Reproductive Biology Research
  - transplantation marker for embryonic and adult tissue
- Toxicology Research
  - B and T cell deficiency, xenograft transplant host
  - xenograft/transplant host
- Developmental Biology Research
  - Cre-lox System
- Genetics Research
  - Mutagenesis and Transgenesis: Cre-lox System
  - Tissue/Cell Markers: Cre-lox System
Immunology, Inflammation and Autoimmunity Research
- Immunodeficiency
  - T cell deficiency
  - B cell defects
  - B cell deficiency
  - B and T cell deficiency
  - specific T cell deficiency
- T Cell Receptor Signaling Defects
  - B and T cell deficiency
  - B and T cell deficiency, xenograft/transplant host
- Autoimmunity
  - B and T cell deficiency
  - B cell deficiency
- Inflammation
  - B and T cell deficiency
- Lymphoid Tissue Defects
  - B and T cell deficiency
  - hematopoietic development
Virology Research
- B and T Cell Deficiency
  - AIDS research tool
Internal/Organ Research
- Thymus Defects
  - B and T cell deficient
- Adrenal Cortex Defects
  - T cell deficiency
- Lymphoid Tissue Defects
  - B and T cell deficiency
  - T cell deficiency
Cancer Research
- Toxicology
  - B and T cell deficiency, xenograft/transplant host
  - xenograft/transplant host
- Cre-lox System
  - loxP-flanked Sequences
Hematological Research
- Immunological Defects
  - B and T cell deficiency
- Hematopoietic Defects
Developmental Biology Research
- Lymphoid Tissue Defects
  - hematopoietic defects
- Internal/Organ Defects
  - hematopoietic defects
  - Lymphoid Tissue Defects
Neurobiology Research
- Cre-lox System
  - loxP-flanked Sequences
  - loxP-flanked Sequences: Test/Reporter

Mammalian Phenotype Terms by Genotype

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

Genotype: Tcf12^tm3Zhu/Tcf12^tm3Zhu Tcf3^tm4Zhu/Tcf3^tm4Zhu Tg(Cd4-cre)1Cwi/0
involves: 129S4/SvJae * C57BL/6 * DBA/2

hematopoietic system phenotype

abnormal CD8-positive, alpha beta T cell morphology
- mice present with CD+TCR- T cell population not found in controls

abnormal T cell differentiation
- T cells prematurely acquire single positive phenotype compared to in wild-type mice

decreased CD4-positive, alpha beta T cell number

increased CD8-positive, alpha-beta T cell number

immune system phenotype

abnormal CD8-positive, alpha beta T cell morphology
- mice present with CD+TCR- T cell population not found in controls

abnormal T cell differentiation
- T cells prematurely acquire single positive phenotype compared to in wild-type mice

decreased CD4-positive, alpha beta T cell number

increased CD8-positive, alpha-beta T cell number

References

Wojciechowski J; Lai A; Kondo M; Zhuang Y. 2007. E2A and HEB are required to block thymocyte proliferation prior to pre-TCR expression. J Immunol 178(9):5717-26PubMed: 17442955 MGI: J:129570
Lazorchak AS; Wojciechowski J; Dai M; Zhuang Y. 2006. E2A promotes the survival of precursor and mature B lymphocytes. J Immunol 177(4):2495-504PubMed: 16888011 MGI: J:138355
Zhang B; Lin YY; Dai M; Zhuang Y. 2014. Id3 and Id2 act as a dual safety mechanism in regulating the development and population size of innate-like gammadelta T cells. J Immunol 192(3):1055-63PubMed: 24379125 MGI: J:207300
Pan L; Hanrahan J; Li J; Hale LP; Zhuang Y. 2002. An Analysis of T Cell Intrinsic Roles of E2A by Conditional Gene Disruption in the Thymus. J Immunol 168(8):3923-32PubMed: 11937548 MGI: J:75746

Additional - Tcf12^tm3Zhu related

Additional - Tcf3^tm4Zhu related

Technical Support

Contact Technical Support

Genotyping Protocols
- Standard PCR:Tcf3^tm4Zhu
- Standard PCR:Tcf12^tm3Zhu
- Genotyping resources and troubleshooting
Breeding Considerations

These HEB^f E2A^f mice harbor two mutations that segregate independently; the HEB floxed allele (Tcf12^tm3Zhu) and E2A floxed allele (Tcf3^tm4Zhu). When maintaining a live colony, mice homozygous for both floxed mutations may be bred together.
- Additional Breeding and Husbandry Support
Citation
When using the HEB^flox E2A^flox mouse strain in a publication, please cite the originating article(s) and include JAX stock #024511 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

Cryo Recovery

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service	Genotype	Price
	Heterozygous for Tcf12<tm3Zhu>, Heterozygous for Tcf3<tm4Zhu>

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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Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.

The Jackson Laboratory's Genotype Promise

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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General Terms and Conditions

Questions about Terms of Use

Licensing Information

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Also Known As: HEBflox E2Aflox

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Approximate Controls

Additional Information

Selected References

Tcf12tm3Zhu

Allele Symbol: Tcf12tm3Zhu

Tcf3tm4Zhu

Allele Symbol: Tcf3tm4Zhu

Disease Terms

Research Areas By Genotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Tcf12tm3Zhu/Tcf12tm3Zhu Tcf3tm4Zhu/Tcf3tm4Zhu Tg(Cd4-cre)1Cwi/0involves: 129S4/SvJae * C57BL/6 * DBA/2

hematopoietic system phenotype

immune system phenotype

References

Additional - Tcf12tm3Zhu related

Additional - Tcf3tm4Zhu related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms of Use

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: HEB^flox E2A^flox

Tcf12^tm3Zhu

Allele Symbol: Tcf12^tm3Zhu

Tcf3^tm4Zhu

Allele Symbol: Tcf3^tm4Zhu

Genotype: Tcf12^tm3Zhu/Tcf12^tm3Zhu Tcf3^tm4Zhu/Tcf3^tm4Zhu Tg(Cd4-cre)1Cwi/0
involves: 129S4/SvJae * C57BL/6 * DBA/2

Additional - Tcf12^tm3Zhu related

Additional - Tcf3^tm4Zhu related