C.Cg-Rag2^tm1Fwa Thy1^a/AkhoAbhmJ

Stock number: 036333
Product name: Thy1.1 RAG-/- Balb/c
Research areas: Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

Thy1.1 RAG-/- Balb/c mice contain a neomycin cassette inserted in the Rag2 (recombination activating gene 2) open reading frame and the Thy1^a (Thy1.1) allelic variant on a BALB background. The addition of the Thy1^a allele to the Rag2 knockout (KO) provides a tracking option for adoptive transfer studies. Rag2 KO mice lack mature T and B cells and are immunodeficient.

Detailed description

Rag2 (recombination activating gene 2) encodes a protein involved in the initiation of V(D)J recombination during B and T cell development. Rag1 and Rag2 form the two core components of the RAG complex; a recombinase responsible for double-stranded DNA breaks at conserved recombination signal sequences. Thy1.1 RAG-/- Balb/c mice contain a neomycin cassette designed to replace a 0.85kb segment of the Rag2 open reading frame, abolishing RAG2 function and the Thy1^a (Thy1.1) allelic variant on a BALB background. Mice homozygous for both alleles are viable, fertile, and normal in size, but should be considered immunodeficient. Rag2 KO mice lack mature T and B cells. These mice may be useful for studying roles of RAG2 in B and T cell development, T and B cell receptor recombination, hematopoiesis, hematology, immunology, and inflammation research. The addition of the Thy1^a allele to the Rag2 knockout (KO) provides a tracking option for adoptive transfer studies and allows for more accurate tracking of transferred cells (either as recipients or donors).

Development

The Rag2 KO was developed by Dr. Frederick Alt at Columbia University. The targeting construct was designed to replace a 0.85kb segment of the open reading frame of the recombination activating gene 2 (Rag2) on chromosome 2 with a neomycin resistance (neo) cassette and a polyadenylation (polyA) sequence. The construct was electroporated into 129S/SvEv-Gpi1^c-derived CCE embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to C57BL/6 females. The strain was maintained on the C57BL/6 background and then backcrossed to the BALB/c background for more than 15 generations. The Thy1^a (Thy1.1) variant (chromosome 9) on a BALB background was then crossed to BALB congenic Rag2 KO in the laboratory of Dr. Alex Khoruts (University of Minnesota). Mice were then transferred to Dr. Clara Abraham who donated them to the The Jackson Laboratory. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize BALB/cJ oocytes (Stock No. 000651).

Allele

Symbol: Rag2^tm1Fwa
Name: targeted mutation 1, Frederick W Alt
MGI accession ID: MGI:1858556
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: RAG-2-; Rag2-; RAG2 KO; Rag2^tm1Alt; Rag2^tm1Fwe; Rag2M
Marker symbol: Rag2
Marker name: recombination activating gene 2
Marker synonyms: Rag-2
Chromosome: 2
Strain of origin: 129S/SvEv-Gpi1^c
Molecular note: A neomycin resistance cassette replaced a 0.85kb segment of the open reading frame, eliminating the encoding of 286 of the 527 amino acids of the protein.

Allele

Symbol: Thy1^a
Name: a variant
MGI accession ID: MGI:3579311
Generation method: Not Applicable
Synonyms: thetaAKR; theta-AKR; Thy1.1; Thy-1.1; Thy1a
Marker symbol: Thy1
Marker name: thymus cell antigen 1, theta
Marker synonyms: CD7; CD90; CDw90; T25; theta; Thy 1.2; Thy-1; Thy1.1; Thy1.2; Thy-1.2
Chromosome: 9
Strain of origin: Not Applicable
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.
Molecular note: The allele Thy1^a determines an antigenic specificity, Thy-1.1, found in the AKR and RF strains.
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).