Rag1-deficient ubiquitin-GFP mice on the C57BL/6 genetic background (B6.Rag1;UBC-GFP) combine lymphocyte-deficiency with widespread green fluorescent protein expression. These mice allow in vivo fluorescent tracking of non-lymphocyte cell populations. They are also suitable for use as recipient hosts for implanted embryos/tissues/cells from non-fluorescent mice.
In 2020, the UBC-GFP transgene was determined to have integrated at Chr17:29,435,589 - a noncoding region centromeric from the H-2 locus (~4.6 Mbp away from H2-K1) - and is closely linked to H-2b MHC haplotype. See Important Note for additional details.
IMR Colony, The Jackson Laboratory
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Null/Knockout) | Rag1 | recombination activating gene 1 |
Allele Type |
---|
Transgenic (Reporter) |
Important Note regarding transgene insertion and linkage to H-2b MHC haplotype: In Liu et al. 2020 J Immunol. 204:1982 [PMID:32122998], it was discovered that the UBC-GFP transgene integrated into chromosome 17 at nucleotide position 29,435,589 (reference sequence NC_000083.6; GRCm38.p6) - this is a noncoding region centromeric from the H-2 locus (~4.6 Mbp away from H2-K1). The number of transgene copies was not determined. Furthermore, this determined that BALB/cBy-congenic UBC-GFP transgenic mice (N11+ from Stock No. 007076) retained the H-2b MHC haplotype from the genetic background upon which it was created (C57BL/6), rather than the H-2d MHC haplotype of BALB/cBy.
Although close to the H-2 locus, segregation is possible. For experiments where MHC haplotype is a factor, researchers should determine if the UBC-GFP transgenic mice are H-2b or H-2d.
In 2020, the UBC-GFP transgene was determined to have integrated at Chr17:29,435,589 - a noncoding region centromeric from the H-2 locus (~4.6 Mbp away from H2-K1) - and is closely linked to H-2b MHC haplotype. See Important Note for additional details.
Rag1-deficient ubiquitin-GFP mice on the C57BL/6 genetic background (B6.Rag1-/-;UBC-GFP) combine lymphocyte-deficiency with widespread green fluorescent protein expression.
Mice homozygous for both the Rag1 null allele and the UBC-GFP transgene (Rag1-/-;UBC-GFP+/+) are viable and fertile. Double homozygous mice produce no mature T cells or B cells, and have widespread GFP expression that is uniform within cell lineage. GFP expression varies between different cell types; hematopoietic cells have high expression levels compared with other cells. Red blood cells from UBC-GFP homozygotes should fluoresce at approximately twice the level compared to UBC-GFP hemizygotes.
Similar to other immunodeficient strains, maintaining Rag1-deficient mice in high health status (specific pathogen-free) vivaria promotes overall colony health. If Rag1-deficient animals are maintained in low health barrier rooms, the use of medicated water (sulfatrim/trimethoprim-sulfa) is suggested to increase overall colony health.
C57BL/6J-congenic mice harboring this Rag1 null allele are described and available from The Jackson Laboratory Repository as Stock No. 002216.
UBC-GFP transgenic mice from founder line 30 on a C57BL/6 genetic background are described and available from The Jackson Laboratory Repository as Stock No. 004353. The transgene insertion on chromosome 17 results in linkage to H-2b MHC haplotype (see Important Note).
These B6.Rag1;UBC-GFP mice harbor the Rag1 null allele (Rag1tm1Mom) and a UBC-GFP transgene (Tg(UBC-GFP)30Scha).
To generate the B6.Rag1;UBC-GFP mutant line (Stock No. 013115), C57BL/6J-congenic Rag1 null homozygous mice (Stock No. 002216) were bred with C57BL/6.UBC-GFP homozygous mice (Stock No. 004353) for three generations and selecting offspring for homozygosity. The Jackson Laboratory Repository will maintain the live Rag1.UBC-GFP colony by breeding mice homozygous for the Rag1 null allele and homozygous for the UBC-GFP transgene.
The Rag1 null allele (Rag1tm1Mom) was designed by Dr. Peter Mombaerts (in the laboratory of Dr. Susumu Tonegawa at the Center for Cancer Research, Massachusetts Institute of Technology) with a PGK-neo cassette replacing 1356 bp (encoding the nuclear localization signal and the zinc-finger motif) at the 5' end of the recombination activating gene 1 coding sequence on chromosome 2. C57BL/6J-congenic mice harboring this Rag1 null allele are described and available from The Jackson Laboratory Repository as Stock No. 002216.
The ubiquitin-GFP transgene (UBI-GFP or UBC-GFP) was designed by Dr. Brian C. Schaefer (while at National Jewish Medical and Research Center) with an enhanced green fluorescent protein open reading frame under the control of the human ubiqutin C promoter. Transgenic mice from founder line 30 on a C57BL/6 genetic background are described and available from The Jackson Laboratory Repository as Stock No. 004353.
In 2020, the UBC-GFP transgene was determined to have integrated at Chr17:29,435,589 - a noncoding region centromeric from the H-2 locus (~4.6 Mbp away from H2-K1) - and is closely linked to H-2b MHC haplotype. See Important Note for additional details.
Expressed Gene | GFP, Green Fluorescent Protein, |
---|---|
Site of Expression | Certain hematopoietic cell types display distinct expression levels of GFP. Expression is uniform within a cell type lineage and remains constant throughout development. |
Depending upon the experiment, the following may also be appropriate control strain(s):
--B6.129S7-Rag1tm1Mom/J (Stock No. 002216)
--C57BL/6-Tg(UBC-GFP)30Scha/J (Stock No. 004353)
--C57BL/6J inbred mice (Stock No. 000664)
Allele Name | targeted mutation 1, Peter Mombaerts |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | Rag-; Rag1tm1Mom; RAG-1-; Rag1-; RAG1null; Rag-1KO |
Gene Symbol and Name | Rag1, recombination activating gene 1 |
Gene Synonym(s) | |
Site of Expression | expression is seen in bone marrow derived cell lines. |
Strain of Origin | 129S7/SvEvBrd-Hprt+ |
Chromosome | 2 |
Molecular Note | A 1356 bp genomic fragment of the Rag1 gene, encoding the nuclear localization signal and the zinc-finger motif, was replaced by a neomycin cassette. A mutant transcript expressed from this allele was detected by Northern blot in bone marrow derived cell lines from homozygous mice. |
Mutations Made By | Peter Mombaerts, Max Planck Research Unit for Neurogenetics |
Allele Name | transgene insertion 30, Brian Schaefer |
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Allele Type | Transgenic (Reporter) |
Allele Synonym(s) | Tg(UBCGFP)30Scha; UBC-GFP; Ub-GFP; UBI-EGFP; UBI-GFP; uGFP |
Gene Symbol and Name | Tg(UBC-GFP)30Scha, transgene insertion 30, Brian Schaefer |
Gene Synonym(s) | |
Promoter | UBC, ubiquitin C, human |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | Certain hematopoietic cell types display distinct expression levels of GFP. Expression is uniform within a cell type lineage and remains constant throughout development. |
Strain of Origin | C57BL/6 |
Chromosome | 17 |
General Note | Homozygous transgenic mice are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. These mice express GFP in all tissues examined. Expression levels vary between certain hematapoietic cell types. GFP expression is uniform within a cell type lineage and remains constant throughout development. T cells have a 2-fold higher GFP expression than CD19+B220+ B cells or peripheral blood cells. Leukocytes and red blood cells from homozygous transgenic mice fluoresce at approximately twice the level of cells from hemizygous mice. |
Molecular Note | The transgene contains an enhanced green fluorescent protein open reading frame under the control of the human ubiquitin C promoter. The transgene integrated into chromosome 17 at nucleotide position 29,435,589 (reference sequence NC_000083.6; GRCm38.p6) - this is a noncoding region centromeric from the H-2 locus (~4.6 Mbp away from H-2 K1). |
Mutations Made By | Brian Schaefer, Uniformed Services Univ. Health Sciences |
The Jackson Laboratory Repository will maintain the live Rag1.UBC-GFP colony by breeding together mice homozygous for both the Rag1 null allele and the UBC-GFP transgene.
Mice homozygous for the Rag1 null allele are lymphocyte-deficient. As such, and similar to other immunodeficient strains, maintenance in high health status (specific pathogen-free) vivaria promotes overall colony health. If Rag1-deficient animals are maintained in low health barrier rooms, the use of medicated water (sulfatrim/trimethoprim-sulfa) is suggested to increase overall colony health.
When using the RAG-1-; UBI-GFP mouse strain in a publication, please cite the originating article(s) and include JAX stock #013115 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Homozygous for Rag1<tm1Mom>, Homozygote for Tg(UBC-GFP)30Scha, |
Frozen Mouse Embryo | B6.Cg-Rag1<tm1Mom> Tg(UBC-GFP)30Scha/J Frozen Embryos | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Rag1<tm1Mom> Tg(UBC-GFP)30Scha/J Frozen Embryos | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Rag1<tm1Mom> Tg(UBC-GFP)30Scha/J Frozen Embryos | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Rag1<tm1Mom> Tg(UBC-GFP)30Scha/J Frozen Embryos | $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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