This b12HL mutant mouse strain expresses the broadly neutralizing HIV antibody b12 and may be useful in design, development and testing of potential HIV vaccines.
David Nemazee, The Scripps Research Institute
Genetic Background | Generation |
---|---|
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Inserted expressed sequence) | Igh | immunoglobulin heavy chain complex |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Inserted expressed sequence) | Igk | immunoglobulin kappa chain complex |
Monoclonal antibodies called broadly neutralizing antibodies, are able to interact with many different HIV isolates of various clades, inhibiting infection of target cells, and are of interest in the design and development of potential protective vaccines. These knock in b12HHLL mice express the broadly neutralizing HIV antibody b12, which recognizes
the CD4 binding site on HIV envelope glycoprotein gp120.
The HIV b12 antibody L and H chain variable exons replace the respective J clusters in the endogenous mouse genes, Igk and Igh. Mice that are heterozygous for the 2 targeted mutations are viable and fertile. More than 90% of B cells from mutant mice heterozygous for the mutations bind soluble HIV envelope antigen as assayed by flow cytometry. Preimmune (naive) sera from heterozygous mutant mice has high titers of HIV Env IgG binding activity.
The Donating Investigator has not attempted to maintain the strain homozygous for both alleles, but reports that double homozygotes should be viable and fertile.
These double mutant mice were generated by crossing the single targeted mutation strains.
For the Igktm1(b12)Nemz allele, b12 L-chain, a targeting vector designed by Dr. David Nemazee (The Scripps Research Institute) containing a floxed NEO cassette was used to replace the J cluster in the Igk locus with the HIV antibody b12 L chain VJ element. The construct was electroporated into C57BL/6NTac-derived iTL IC1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were then bred to Cre recombinase expressing mice on the congenic C57BL/6J background (Stock No. 003724) to remove the floxed NEO cassette.
For the Ightm1(b12)Nemz allele, b12 H-chain, a targeting vector designed by Dr. David Nemazee (The Scripps Research Institute) containing a floxed NEO cassette was used to replace the DQ52-JH cluster in the Igh locus with HIV antibody b12 variable region coding sequence. The construct was electroporated into C57Bl/6-derived C2 ES cell line C57Bl/6-derived C2 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were then bred to Cre recombinase expressing mice on the congenic C57BL/6J background (Stock No. 003724) to remove the floxed NEO cassette.
The single targeted mutation strains were crossed to generate the double mutant line (double heterozygotes). Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
Allele Name | targeted mutation 1.1, David Nemazee |
---|---|
Allele Type | Targeted (Inserted expressed sequence) |
Allele Synonym(s) | b10H |
Gene Symbol and Name | Igh, immunoglobulin heavy chain complex |
Gene Synonym(s) | |
Strain of Origin | C57BL/6NCrl |
Chromosome | 12 |
Molecular Note | A floxed neomycin resistance cassette replaced the DQ52-JH cluster in the Igh locus with HIV antibody b12 variable region coding sequence. Cre-mediated recombination removed the neomycin resistance cassette. |
Allele Name | targeted mutation 1.1, David Nemazee |
---|---|
Allele Type | Targeted (Inserted expressed sequence) |
Allele Synonym(s) | b10L |
Gene Symbol and Name | Igk, immunoglobulin kappa chain complex |
Gene Synonym(s) | |
Strain of Origin | C57BL/6NTac |
Chromosome | 6 |
Molecular Note | A floxed neomycin resistance cassette replace the J cluster in the Igk locus with the HIV antibody b12 L chain VJ element. Cre-mediated recombination removed the neomycin resistance cassette. |
When maintaining a live colony, these mice can be bred as heterozygotes for both alleles. The Donating Investigator has not attempted to maintain the strain homozygous for both alleles, but reports that double homozygotes should be viable and fertile.
When using the C57BL/6(FVB)-Igktm1.1(b12)Nemz Ightm1.1(b12)Nemz/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #023065 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Igk<tm1.1(b12)Nemz> , Heterozygous or wildtype for Igh<tm1.1(b12)Nemz> |
Frozen Mouse Embryo | C57BL/6(FVB)-Igk<tm1.1(b12)Nemz> Igh<tm1.1(b12)Nemz>/J | $2595.00 |
Frozen Mouse Embryo | C57BL/6(FVB)-Igk<tm1.1(b12)Nemz> Igh<tm1.1(b12)Nemz>/J | $2595.00 |
Frozen Mouse Embryo | C57BL/6(FVB)-Igk<tm1.1(b12)Nemz> Igh<tm1.1(b12)Nemz>/J | $3373.50 |
Frozen Mouse Embryo | C57BL/6(FVB)-Igk<tm1.1(b12)Nemz> Igh<tm1.1(b12)Nemz>/J | $3373.50 |
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