Overview

Also Known As:Bmi-1GFP

The Bmi-1^GFP knock-in/knock-out allele was designed to both abolish Bmi1 gene function and express EGFP from the Bmi1 promoter/enhancer elements. Of note, these mice also harbor the CD45.2 (Ptprc^b) and Thy1.1 leukocyte alloantigen (Thy1^a) alleles to allow tracking of hematopoietic donor derived or transferred cells.

Donating Investigator

Irving L Weissman, Stanford University

Genetic overview

Genetic Background	Generation

Bmi1^tm1Ilw

Allele Type	Gene Symbol	Gene Name
Targeted (Reporter, Null/Knockout)	Bmi1	Bmi1 polycomb ring finger oncogene

Ptprc^b

Allele Type	Gene Symbol	Gene Name
Not Applicable	Ptprc	protein tyrosine phosphatase, receptor type, C

Thy1^a

Allele Type	Gene Symbol	Gene Name
Not Applicable	Thy1	thymus cell antigen 1, theta

View Genetics

Research Applications

Research Tools
Developmental Biology Research
Immunology, Inflammation and Autoimmunity Research
Cell Biology Research
Hematological Research

View All Research Applications

Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

The Bmi-1^GFP knock-in/knockout allele has EGFP fused in-frame with the endogenous transcriptional start site of the Bmi-1 polycomb ring finger oncogene (Bmi1) locus; both abolishing endogenous gene function and placing EGFP expression under direction of the Bmi1 promoter/enhancer regions. EGFP expression (direct fluorescence) is highest in hematopoietic stem cell (HSC) populations and downregulated along with differentiation; recapitulating the expected expression pattern of endogenous Bmi-1. Specifically, EGFP fluorescence is significantly higher in the HSC-enriched KLS population (c-kit⁺lin^-Sca-1⁺) compared with other populations.

Mice heterozygous for the Bmi-1^GFP knock-in/knockout allele are viable, fertile, and phenotypically indistinguishable from wildtype littermate controls with respect to survival, hematopoietic cellularity, and lineage composition. Heterozygous bone marrow cells sorted via FACS into GFP^high and GFP^lo show that the Bmi-1 expression level in the GFP^high population is approximately 10-fold higher than that of the GFP^lo population; these Bmi-1^high cell populations are functional HSCs.

The donating investigator reports that mice homozygous for the Bmi-1^GFP knock-in/knockout allele die before birth or soon after they are born. In addition, homozygotes may be expected to exhibit the same phenotype as mice homozygous for other null mutations of this gene (decreased hematopoietic cell number, immune deficiency, neurological abnormalities, and posterior transformation).

These mice also harbor the Ptprc^b allele, CD45.2, of C57BL/6J origin rather than the alternate allele normally present in C57BL/Ka inbred mice. As CD45 is expressed on all adaptive and innate immune cells, this strain provides a unique cell surface marker that can be used to track lineage and phenotype of all hematopoietic donor derived or transferred cells.

In addition, these mice also harbor the Thy1.1 leukocyte alloantigen allele, Thy1^a, rather than the alternate allele normally present in C57BL/Ka inbred mice (as well as C57BL/6, C57BL/10, DBA/2, NOD, and BALB/c inbred mice). Thus, cell populations derived from these mutant mice can be distinguished from syngeneic host and other mice with the alternate allele via flow cytometry. The presence of Thy1^a serves as a marker for tracking donor cells in vitro.

Development

A targeting vector was designed to replace exon 2 of the Bmi1 polycomb ring finger oncogene (Bmi1) locus with an enhanced green fluorescent protein (EGFP) and loxP-flanked neomycin resistance cassette. The EGFP is inserted in-frame to the Bmi1 ATG translation start site. The construct was electroporated into (129X1/SvJ x 129S1/Sv)F1-Kitl⁺)-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were then transiently transfected with a Cre recombinase-expressing plasmid. The resulting ES cells with the Bmi-1^GFP genotype (neo selection cassette removed; leaving EGFP and a single loxP site replacing exon 2) were injected into recipient blastocysts. Chimeric animals were bred to C57BL/Ka mice with the CD45.2 and Thy1.1 alleles (C57BL/Ka-Ptprc^b Thy1^a) to generate the colony. The Bmi-1^GFP mutant mice were backcrossed to the C57BL/Ka-CD45.2::Thy1.1 line for more than twenty generations. C57BL/Ka-CD45.2::Thy1.1 females and C57BL/Ka-Bmi-1^GFP::CD45.2::Thy1.1 males were sent to The Jackson Laboratory Repository. Upon arrival, these mice were bred together to establish the colony.

Expression Data

Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	EGFP expression (direct fluorescence) is highest in hematopoietic stem cell (HSC) populations and downregulated along with differentiation.
Site of Expression	Widely expressed on all adaptive and innate immune cells.
Site of Expression	The Thy1 locus determines a surface antigen present on cells of the thymus, a number of mouse leukemias, brain, and in lesser amounts on lymph node and spleen cells.

Control Suggestions

The appropriate control mouse are mice from the colony that are wildtype at the Bmi1 locus and homozygous for both the Ptprc^b and Thy1^a alleles. This control mouse is expected to be synonymous with the C57B/Ka-Ptprc^b Thy1^a mice used by the donating investigator when backcrossing Stock No. 017351.

Additional Information

Considerations for Choosing Controls

Genetics

Bmi1^tm1Ilw

Allele Symbol: Bmi1^tm1Ilw

Allele Name	targeted mutation 1, Irving L Weissman
Allele Type	Targeted (Reporter, Null/Knockout)
Allele Synonym(s)	Bmi-1^GFP; Bmi1^tm1Nhsn
Gene Symbol and Name	Bmi1, Bmi1 polycomb ring finger oncogene
Gene Synonym(s)
Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	EGFP expression (direct fluorescence) is highest in hematopoietic stem cell (HSC) populations and downregulated along with differentiation.
Strain of Origin	(129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Chromosome	2
Molecular Note	Exon 2 was replaced with an in frame EGFP cassette, exon 2 and floxed neo cassette. Transient cre expression in ES cells was used to remove the neo cassette. Expression of the EGFP precludes expression of the endogenous gene.
Mutations Made By	Irving Weissman, Stanford University

Ptprc^b

Allele Symbol: Ptprc^b

Allele Name	b variant
Allele Type	Not Applicable
Allele Synonym(s)	CD45.2; Ly5^b
Gene Symbol and Name	Ptprc, protein tyrosine phosphatase, receptor type, C
Gene Synonym(s)
Site of Expression	Widely expressed on all adaptive and innate immune cells.
Strain of Origin	Not Applicable
Chromosome	1
Molecular Note	Ptprc^b is found in strains C57BL/6, C3H/An, DBA/2, AKR, and many others (J:13367, J:12054, J:12077, J:8603). Twelve nucleotide differences between the a and b alleles have been identified. These base substitutions correspond to five amino-acid changes within the extracellular domain of the encoded protein. These amino-acid differences are clustered in a region that also contains the greatest divergence between mouse and rat sequences (J:22485). Note that the allele designations originally described were reversed in 1987 (J:8603); all publications prior to 1987 show SJL/J, STS/A, and DA as having the b allele and the C57BL/6J group as having the a allele (J:22341).

Thy1^a

Allele Symbol: Thy1^a

Allele Name	a variant
Allele Type	Not Applicable
Allele Synonym(s)	thetaAKR; theta-AKR; Thy1.1; Thy-1.1; Thy1a
Gene Symbol and Name	Thy1, thymus cell antigen 1, theta
Gene Synonym(s)
Site of Expression	The Thy1 locus determines a surface antigen present on cells of the thymus, a number of mouse leukemias, brain, and in lesser amounts on lymph node and spleen cells.
Strain of Origin	Not Applicable
Chromosome	9
General Note	The Thy1 locus determines a surface antigen present on cells of the thymus, a number of mouse leukemias, brain, and in lesser amounts on lymph node and spleen cells. The allele Thy1^a determines an antigenic specificity, Thy-1.1, found in the AKR and RF strains; the allele Thy1^b determines an antigenic specificity, Thy-1.2, found in the C3HeB/Fe and many other strains (J:5243, J:5012, J:4469). The Thy1 antigen is probably present on all T lymphocytes and absent from all B lymphocytes, and it thus serves as a valuable T-cell marker (J:5243). It is very widely used in experiments designed to determine the distribution and function of T-cells. Thy1 specifies a T-cell surface glycoprotein, T25, with a molecular weight of 25 kDa (J:5707). The protein appears to be anchored in the cell membrane by a lipid that is either phosphotidylinositol or closely related to it (J:12016). Thy1 may function in the cell membrane as a signal transduction molecule (J:8333). The Thy1 locus, or possibly a gene closely linked to it, controls quantitative expression of a protein that is the same size as Thy1 and is expressed on thymus and brain but not on lymph node and spleen cells (J:7900).
Molecular Note	The allele Thy1^a determines an antigenic specificity, Thy-1.1, found in the AKR and RF strains.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Research Tools
- Fluorescent Proteins
- Hematological Research
- Genetics Research
  - Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
  - Tissue/Cell Markers: cell marker for bone marrow transplantation
  - Tissue/Cell Markers
- Reproductive Biology Research
  - transplantation marker for embryonic and adult tissue
- Developmental Biology Research
  - transplantation marker for embryonic and adult tissue
- Cell Biology Research
Developmental Biology Research
- Embryonic Lethality (Homozygous)
  - incomplete
- Lymphoid Tissue Defects
  - hematopoietic defects
- Internal/Organ Defects
  - hematopoietic defects
Cell Biology Research
- Cell Cycle Regulation
Hematological Research
- Hematopoietic Defects

Genotype: Thy1^a related

Immunology, Inflammation and Autoimmunity Research
- CD Antigens, Antigen Receptors, and Histocompatibility Markers
Research Tools
- Immunology, Inflammation and Autoimmunity Research
  - T cell specific surface marker
- Genetics Research
  - Tissue/Cell Markers: T cell specific surface marker
  - Tissue/Cell Markers

Mammalian Phenotype Terms by Genotype

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

Genotype: Bmi1^tm1Ilw/Bmi1⁺
involves: 129S1/Sv * 129X1/SvJ * C57BL/6

normal phenotype

no abnormal phenotype detected
- Normal - mice are indistinguishable from wild-type mice

References

Additional - Bmi1^tm1Ilw related

Additional - Ptprc^b related

Additional - Thy1^a related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Probe:Ptprc End Point-Alternate 1

Breeding Considerations

When maintaining a live colony, mice heterozygotes for the Bmi1^GFP mutation and homozygous for both the Ptprc^b and Thy1^a alleles are bred with mice from the colony that are wildtype at the Bmi1 locus and homozygous for both the Ptprc^b and Thy1^a alleles. Mice homozygous for the Bmi1^GFP mutation die before birth or soon after they are born.

Additional Breeding and Husbandry Support

Citation
When using the Bmi-1GFP mouse strain in a publication, please cite the originating article(s) and include JAX stock #017351 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

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International

Live Mouse

Age Genotype Price

weeks

Cryorecovery - Pricing

Service/Product Description Price

Homozyogus for Ptprc,Heterozygous or wildtype for Bmi1<tm1Ilw>, Homozygous for Thy1<a>

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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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Also Known As:Bmi-1GFP

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Bmi1tm1Ilw

Allele Symbol: Bmi1tm1Ilw

Ptprcb

Allele Symbol: Ptprcb

Thy1a

Allele Symbol: Thy1a

Disease Terms

Research Areas By Phenotype

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

Genotype: Thy1a related

Mammalian Phenotype Terms by Genotype

Genotype: Bmi1tm1Ilw/Bmi1+involves: 129S1/Sv * 129X1/SvJ * C57BL/6

normal phenotype

References

Additional - Bmi1tm1Ilw related

Additional - Ptprcb related

Additional - Thy1a 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

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

Bmi1^tm1Ilw

Allele Symbol: Bmi1^tm1Ilw

Ptprc^b

Allele Symbol: Ptprc^b

Thy1^a

Allele Symbol: Thy1^a

Genotype: Thy1^a related

Genotype: Bmi1^tm1Ilw/Bmi1⁺
involves: 129S1/Sv * 129X1/SvJ * C57BL/6

Additional - Bmi1^tm1Ilw related

Additional - Ptprc^b related

Additional - Thy1^a related