This double targeted mutation of the immunoglobulin kappa chain complex (Igk) and immunoglobulin heavy chain complex (Igh) genes displays an almost monoclonal B cell population and may be useful in studies of B cell development, receptor editing, tolerance and antibody diversification.
Klaus Rajewsky, Max Delbruck Centre for Molecular Medicine
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Inserted expressed sequence) | Igk | immunoglobulin kappa chain complex |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted | Igh | immunoglobulin heavy chain complex |
Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. As expected, most splenic B cells express the 3-83 idiotype, but total B cell number may be lower than wild-type. The 3-83 antibody is non-autoreactive on the H-2d background, while it is autoreactive on the H-2b and H-2k backgrounds. Mice heterozygous for both mutations and/or the H-2b and H-2k backgrounds exhibit an increase in frequency and absolute number of lambda-expressing B lymphocytes and a decrease in the expression of the 3-83 idiotype due to Igk RS recombination and Vl to Jl rearrangements. The 3-83KiJk<-> allele lacks endogenous Jk segments, preventing secondary Vk to Jk rearrangements.
Mice homozygous for both mutations on the H-2d, but not H-2b and H-2k backgrounds, do not retain 3-83 expression and do not have Vk to Jk recombination events.
This strain generates an almost monoclonal B cell population. This mutant mouse strain may be useful in studies of B cell development, receptor editing, tolerance and antibody diversification.
The targeting at the Igh locus was performed to replace the DNA region from upstream of DQ52 to downstream of JH4 with a vector that contains a loxP-flanked neo cassette, followed by the Ig 3-83 heavy chain gene, which includes its promoter region and the rearranged VDJ (with JH1). The construct was electroporated into 129P2/OlaHsd-derived E14.1TG3B1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The neomycin cassette was removed by transient transfection with a Cre recombinase expressing plasmid leaving a single loxP site. The resulting chimeric animals were crossed to C57BL/6 mice.
The targeting at the Igk locus was performed to replace the DNA region from upstream of Jk1 to downstream of Jk5 with a vector that contains a loxP-flanked neo cassette, followed by the Ig 3-83 light (kappa-k) chain gene, which includes its promoter region and the rearranged VJ (with Jk2), removing all endogenous Jk segments and preventing secondary Vk-Jk rearrangements. The construct was electroporated into 129P2/OlaHsd-derived E14.1TG3B1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The neomycin cassette was removed by transient transfection with a Cre recombinase expressing plasmid leaving a single loxP site 5' to the Vk gene. The resulting chimeric animals were crossed to C57BL/6 mice.
Mice carrying the two mutations were bred together and then backcrossed to BALB/c (H2d) for 10 generations.
Allele Name | targeted mutation 2, Klaus Rajewsky |
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Allele Type | Targeted (Inserted expressed sequence) |
Allele Synonym(s) | 3-83kappaiJkappa-; Igktm2Pld |
Gene Symbol and Name | Igk, immunoglobulin kappa chain complex |
Gene Synonym(s) | |
Promoter | Igk, immunoglobulin kappa chain complex, mouse, laboratory |
Strain of Origin | Not Specified |
Chromosome | 6 |
Molecular Note | A targeting vector was used to insert a recombined Vk3-83-Jk5 region where all endogenous Jkappa segments are deleted. |
Allele Name | targeted mutation 2, Klaus Rajewsky |
---|---|
Allele Type | Targeted |
Allele Synonym(s) | 3-83Hi; 3-83mCkappa; Ightm1Pld |
Gene Symbol and Name | Igh, immunoglobulin heavy chain complex |
Gene Synonym(s) | |
Promoter | Igh, immunoglobulin heavy chain complex, mouse, laboratory |
Strain of Origin | 129P2/OlaHsd |
Chromosome | 12 |
Molecular Note | The rearranged 3-83 heavy chain was inserted into the IgHa locus via homologous recombination. A floxed neo selection cassette was also introduced and subsequently removed by Cre mediated recombination. |
While maintaining a live colony, these mice are bred as homozygotes for both mutations.
When using the C.129P2(B6)-Igktm2Rsky Ightm2Rsky/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #012235 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Igk<tm2Rsky>, Heterozygous for Igh<tm2Rsky |
Frozen Mouse Embryo | C.129P2(B6)-Igk<tm2Rsky> Igh<tm2Rsky>/J | $2595.00 |
Frozen Mouse Embryo | C.129P2(B6)-Igk<tm2Rsky> Igh<tm2Rsky>/J | $2595.00 |
Frozen Mouse Embryo | C.129P2(B6)-Igk<tm2Rsky> Igh<tm2Rsky>/J | $3373.50 |
Frozen Mouse Embryo | C.129P2(B6)-Igk<tm2Rsky> Igh<tm2Rsky>/J | $3373.50 |
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