B6.Cg-Rag1^tm1Mom Tg(UBC-GFP)30Scha/J

Stock number: 013115
Product name: RAG-1^-; UBI-GFP
Research areas: Cancer Research, Diabetes and Obesity Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Neurobiology Research, Research Tools, Virology Research

Description

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-2^b MHC haplotype. See Important Note for additional details.

Detailed description

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-2^b 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-2^b MHC haplotype (see Important Note).

Development

These B6.Rag1;UBC-GFP mice harbor the Rag1 null allele (Rag1^tm1Mom) 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 (Rag1^tm1Mom) 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-2^b MHC haplotype. See Important Note for additional details.



In 2018, a 48 SNP (single nucleotide polymorphism) panel analysis, with 43 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 rederived living colony at The Jackson Laboratory Repository. Three of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating, including the rd8 recessive mutation associated with retinal degeneration on Chromosome 1 within the Crb1 locus. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Tg(UBC-GFP)30Scha
Name: transgene insertion 30, Brian Schaefer
MGI accession ID: MGI:3057178
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(UBC-GFP)30Scha
Marker name: transgene insertion 30, Brian Schaefer
Chromosome: 17
Strain of origin: C57BL/6
Expressed genes: 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.
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).
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.

Allele

Symbol: Rag1^tm1Mom
Name: targeted mutation 1, Peter Mombaerts
MGI accession ID: MGI:1857241
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Rag1
Marker name: recombination activating gene 1
Chromosome: 2
Strain of origin: 129S7/SvEvBrd-Hprt⁺
Site of expression: expression is seen in bone marrow derived cell lines.
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.