An internal ribosome entry site (IRES)-enhanced green fluorescent protein (eGFP) fusion protein was placed downstream of exon 5 of the interleukin 10 (Il10) gene. GFP is expressed in B cells, T cells, myeloid cells, dendritic cells, and Natural Killer cells. These mice may be useful as a tool for studying Il10 gene regulation.
Christopher Karp, Bill & Melinda Gates Foundation
Genetic Background | Generation |
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F?+pN2F6
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Reporter) | Il10 | interleukin 10 |
A targeting vector was designed to place an internal ribosome entry site (IRES)-enhanced green fluorescent protein (eGFP) fusion protein, downstream of exon 5 of the interleukin 10 (Il10) gene. Homozygous mice are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. Il10 is expressed in leukocytes and other cells and is integral to immune homeostasis. These Vert-X mice exhibit baseline intracellular GFP expression in 1.3% of leukocytes. Specifically, GFP expression was seen in B cells, T cells, myeloid cells, dendritic cells, and Natural Killer (NK) cells. B cells were the predominant expressers of GFP with plasma cells, plasmablasts, and CD19+B220+ cells being the highest expressing. GFP expression was seen in an increased amount of leukocytes after challenge with LPS. These mice may be useful as a tool for studying the localization and initiation of Il10 gene regulation.
A targeting vector was designed to place a loxP-flanked neomycin (neo) resistance cassette, and an internal ribosome entry site (IRES)-enhanced green fluorescent protein (eGFP) fusion protein, downstream of exon 5 of the interleukin 10 (Il10) gene. The construct was electroporated into C57BL/6-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into albino C57BL/6 blastocysts and the resulting chimeric males were bred to female albino C57BL/6 mice. Offspring, heterozygous for this floxed allele, were bred with mice expressing Zp3-Cre recombinase on a C57BL/6 background to delete the neo cassette, and progeny were crossed resulting in a colony of Vert-X mice. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation to establish the colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
---|---|
Expressed Gene | Il10, interleukin 10, mouse, laboratory |
Site of Expression | GFP and Il10 are expressed in activated homozygous B cells. |
Allele Name | targeted mutation 1.1, Christopher L Karp |
---|---|
Allele Type | Targeted (Reporter) |
Allele Synonym(s) | Il10gfp; Vert-X |
Gene Symbol and Name | Il10, interleukin 10 |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | Il10, interleukin 10, mouse, laboratory |
Site of Expression | GFP and Il10 are expressed in activated homozygous B cells. |
Strain of Origin | C57BL/6 |
Chromosome | 1 |
Molecular Note | A floxed neomycin cassette and an IRES- eGFP cassette was cloned into the HindIII site between the endogenous stop and poly(A) sites of the gene locus. The neomycin cassette was subsequently removed by breeding founder mice with cre-transgenic mice. The targeted locus expressed both the endogenous gene and the reporter gene in activated homozygous B cells as determined by ELISPOT analysis on GFP+ cells. |
Mutations Made By | Christopher Karp, Bill & Melinda Gates Foundation |
When maintaining a live colony, homozygous mice may be bred together.
When using the Vert-X (or VertX) mouse strain in a publication, please cite the originating article(s) and include JAX stock #014530 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Heterozygous for Il10<tm1.1Karp> |
Frozen Mouse Embryo | B6(Cg)-Il10<tm1.1Karp>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6(Cg)-Il10<tm1.1Karp>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6(Cg)-Il10<tm1.1Karp>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6(Cg)-Il10<tm1.1Karp>/J Frozen Embryo | $3373.50 |
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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