B6(Cg)-Il10^tm1.1Karp/J

Stock number: 014530
Product name: Vert-X (or VertX)
Strain synonyms: Il10^gfp
Research areas: Cell Biology Research, Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

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.

Detailed description

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.

Development

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.

Allele

Symbol: Il10^tm1.1Karp
Name: targeted mutation 1.1, Christopher L Karp
MGI accession ID: MGI:4355198
Generation method: Targeted
Attributes: Reporter
Marker symbol: Il10
Marker name: interleukin 10
Chromosome: 1
Strain of origin: C57BL/6
Expressed genes: GFP,Green Fluorescent Protein,; Il10,interleukin 10,mouse, laboratory
Site of expression: GFP and Il10 are expressed in activated homozygous B cells.
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.