These "kappa-macroself Ag#2" transgenic mice may be useful to study B cell receptor editing, B cell tolerance in polyclonal immune systems, and for the rapid screening of mutants of interest for potential defects in B cell tolerance.
David Nemazee, The Scripps Research Institute
Genetic Background | Generation |
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|
Allele Type |
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Transgenic (Inserted expressed sequence) |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Not Applicable | Ptprc | protein tyrosine phosphatase, receptor type, C |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Not Applicable | Pepc | peptidase C |
"Kappa-macroself Ag#2" transgenic mice express a membrane-bound "macroself" superantigen with specificity for the mouse immunoglobulin kappa light chain (IgΚ). Hemizygous mice are viable and fertile. The transgene is expressed in virtually all cells tested, including lymphoid tissue and bone marrow. B cells expressing the IgΚ-macroself superantigen are absent from the peripheral lymphoid organs while Igl+ B cell numbers are substantially increased. Although bone marrow B cell numbers are unchanged, total peripheral B cell populations are reduced by approximately one half. These mice also express the CD45.1 (Ly5.1 or Ptprca) allele, which is atypical for this C57BL/6 (B6) congenic background, and this marker may be used to track donor cell populations in bone marrow transplantation studies with B6 (CD45.2: Ptprcb) mice.
These "kappa-macroself Ag#2" transgenic mice may be useful to study B cell receptor editing and B cell tolerance in a polyclonal immune system, and to rapidly screen other mutations for potential defects in B cell tolerance.
Genotyping Note: As the transgene expresses a superantigen reactive to immunoglobulin kappa of mouse and may interfere with antibody specificity if using mouse anti-CD45 antibodies, a pre-blocking step using unlabeled mouse IgG:kappa (or equivalent blocking step) is recommended for FACS-typing of the Ptprc (CD45) allele.
These mice are designed to express a membrane-tethered single-chain variable fragment antibody with specificity to mouse IgΚ light chain. The 4 kb Ulik transgene contains the 1.2 kb human ubiquitin C promoter, rheumatoid factor Ig light chain gene leader exon and first intron, the expressed variable genes from the rat anti-mouse Ig CΚ hybridoma 187 line, the hinge, CH2, and CH3 sequences from rat IgG1 heavy chain cDNA, and the transmembrane and intracytoplasmic sequences from the mouse H2-kb gene. This construct was microinjected into (B6xDBA/2)F1 embryos and then implanted into pseudopregnant Igk-deficient foster dams. "Kappa-macroself" transgenic mice (founder line 2) were backcrossed to mice on a "B6.CD45.1" background (derived from Stock No. 002014, B6.SJL-Ptprca Pepcb/BoyJ) for approximately 20 generations before arriving at The Jackson Laboratory.
Expressed Gene | scFv, single chain variable fragment, |
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Site of Expression | |
Site of Expression | Widely expressed on all adaptive and innate immune cells. |
Site of Expression |
Allele Name | transgene insertion 2, David Nemazee |
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Allele Type | Transgenic (Inserted expressed sequence) |
Allele Synonym(s) | kappa-macroself; Ulik |
Gene Symbol and Name | Tg(UBC-scFv)2Nemz, transgene insertion 2, David Nemazee |
Gene Synonym(s) | |
Promoter | UBC, ubiquitin C, human |
Expressed Gene | scFv, single chain variable fragment, |
Strain of Origin | (C57BL/6 x DBA/2)F1 |
Chromosome | UN |
Molecular Note | These mice are designed to express a membrane-tethered single chain variable fragment antibody with specificity to mouse Igk light chain. The 4 kb "Ulik" transgene contained the 1.2 kb human ubiquitin C promoter, rheumatoid factor Ig light chain gene leader exon and first intron, the expressed variable genes from the rat anti-k hybridoma 187 line, the hinge, CH2, and CH3 sequences from rat gamma chain 1 (CH1 domain deleted), and the transmembrane and intracytoplasmic sequences from the H2-kb gene. |
Mutations Made By | Djemel Ait Azzouzene , The Scripps Research Institute |
Allele Name | a variant |
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Allele Type | Not Applicable |
Allele Synonym(s) | CD45.1; Ly5a; PtprcSJL |
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 | Ptprca is found in strains SJL/J, STS/A, and DA. Ptprcb 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). |
Allele Name | b variant |
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Allele Type | Not Applicable |
Allele Synonym(s) | Pep3b |
Gene Symbol and Name | Pepc, peptidase C |
Gene Synonym(s) | |
Strain of Origin | Not Applicable |
Chromosome | 1 |
Molecular Note | The Pepc gene controls electrophoretic variation of peptidase C (PEP-C; EC 3.4.11.-) of red blood cells. The allele Pepcb determines a faster migrating band and is found in the CBA/J and A/He strains. |
When maintaining a live colony, hemizygous mice can be bred to wildtype siblings or to Stock No. 002014. The donating investigator has not attempted to breed hemizygous mice together.
Genotyping Note: As the transgene expresses a superantigen reactive to immunoglobulin kappa of mouse and may interfere with antibody specificity if using mouse anti-CD45 antibodies, a pre-blocking step using unlabeled mouse IgG:kappa (or equivalent blocking step) is recommended for FACS-typing of the Ptprc (CD45) allele.
When using the B6.Cg-Pepcb Ptprca Tg(UBC-scFv)2Nemz/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #006259 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Homozygous for Pepc<b>, Homozygous for Ptprc<a>, Hemizygous or Non carrier for Tg(UBC-scFv)2Nemz |
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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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