A membrane-bound form of enhanced green fluorescent protein (EGFP) replaces the coding region of Clec9a and abolishes gene expression.
Caetano Reis e Sousa, London Research Institute
Genetic Background | Generation |
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N7+pN1F17
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter, Null/Knockout) | Clec9a | C-type lectin domain family 9, member a |
The coding region of the C-type lectin domain family 9, member a (Clec9a) gene is replaced by a membrane-bound farnesylated form of enhanced green fluorescent protein (EGFP), abolishing gene expression. Clec9a encodes for DNGR-1, an endocytic receptor of the C-type lectin superfamily that is selectively expressed at high levels by CD8α+ dendritic cells (DC) and at lower levels by plasmacytoid DC. DNGR-1 recognizes an intracellular ligand that is exposed in dead cells and facilitates cross-presentation of dead-cell-associated antigens through a mechanism that involves interaction of its intracellular tail with the tyrosine kinase, SYK. Homozygous mice are viable and fertile. In these mice, EGFP expression is detected on CD8α+ DC but not on plasmacytoid DC.
DNGR-1 KO mice exhibit a selective deficiency in cross-priming of CD8+ T cells against dead-cell-associated antigens. The phagocytosis of dead cells by DCs is unaffected by loss of DNGR-1 and DC number and phenotype are also unchanged compared to control. These mice may be useful for studying adaptive immune responses to antigens in apoptotic and necrotic cells.
A targeting vector was designed to replace the C-type lectin domain family 9, member a (Clec9a) coding region with a farnesylated form of enhanced green fluorescent protein (EGFP), a polyadenylation signal, and a loxP-flanked neomycin (neo) resistance cassette. This construct was electroporated into (129S6/SvEvTac x C57BL/6) F1-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient C57BL/6 blastocysts and chimeric males were bred with C57BL/6J females. Mutant mice were bred with Tg(Pgk-cre)1Lni mice (on a C57BL/6J congenic background) to remove the neo selection cassette. These Clec9agfp/gfp mice were subsequently bred to C57BL/6J mice for at least 7 generations. Upon arrival at The Jackson Laboratory Repository, mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Site of Expression | EGFP expression is detected on CD8α+ dendritic cells but not on plasmacytoid DC. |
Allele Name | targeted mutation 1.1, Caetano Reis e Sousa |
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Allele Type | Targeted (Reporter, Null/Knockout) |
Allele Synonym(s) | Clec9agfp |
Gene Symbol and Name | Clec9a, C-type lectin domain family 9, member a |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | EGFP expression is detected on CD8α+ dendritic cells but not on plasmacytoid DC. |
Strain of Origin | (129S6/SvEvTac x C57BL/6J)F1 |
Chromosome | 6 |
General Note | The expression of NK1.1 (Klrb1c) is linked to Clec9a deficiency in homozygous mice, indicating that the homologous recombination step targeted the chromosome of C57BL/6J origin. |
Molecular Note | A targeting vector was designed to replace the C-type lectin domain family 9, member a (Clec9a) coding region with a farnesylated form of enhanced green fluorescent protein (EGFP), a polyadenylation signal, and a loxP-flanked neomycin (neo) resistance cassette. Cre-mediated recombination removed the neo cassette. |
Mutations Made By | Caetano Reis e Sousa, London Research Institute |
When maintaining a live colony, homozygous mice may be bred together.
When using the Clec9agfp mouse strain in a publication, please cite the originating article(s) and include JAX stock #017696 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Clec9a<tm1.1Crs> |
Frozen Mouse Embryo | B6(Cg)-Clec9a<tm1.1Crs>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6(Cg)-Clec9a<tm1.1Crs>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6(Cg)-Clec9a<tm1.1Crs>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6(Cg)-Clec9a<tm1.1Crs>/J Frozen Embryo | $3373.50 |
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