C57BL/6-Il17a^tm1Bcgen/J

Stock number: 018472
Product name: IL17A-IRES-GFP-KI
Research areas: Research Tools

Description

These IL17-GFP mice express EGFP as a marker of Il17a activity. Fluorescence can be observed in some intraepithelial lymphocytes (IELs) of the small intestine without stimulation. Naive CD4⁺ T cells can be stimulated to show increased fluorescence associated with IL17A production.

Detailed description

IL17A (interleukin 17A; also known as CTLA8) is a pro-inflammatory cytokine that, although expressed by several cell types, defines the T helper 17 (T^H17) cell subset. IL17A activates signal transduction pathways that recruit neutrophils, promote the production of antimicrobial factors, and drive the inflammatory pathology associated with autoimmune disorders.

These mice express enhanced green fluorescent protein (EGFP) as a marker of Il17a activity. Fluorescence can be observed in some intraepithelial lymphocytes (IELs) of the small intestine without stimulation. Naive CD4⁺ T cells stimulated with plate-bound anti-CD3/anti-CD28 in the presence of transforming growth factor, beta 1 (TGFB, 1 ng/ml) and interleukin 6 (IL6, 20 ng/ml) show increased fluorescence associated with IL17A production. Normal levels of IL17A are expressed.

Please follow this link for more in vivo and in vitro phenotype data.

Development

An IRES-EGFP-SV40-polyA signal sequence cassette was inserted after the stop codon of the Il17a gene using C57BL/6-derived embryonic stem (ES) cells. This strain was maintained on a C57BL/6 genetic background by the donating laboratory prior to sending to The Jackson Laboratory. Upon arrival, sperm was cryopreserved. To generate the living colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J female mice (Stock No. 000664). The mutant mice were then bred together to maintain the colony.

In December of 2020 we learned that a loxP-flanked neomycin cassette is located just after the GFP coding region/3' UTR. The neomycin cassette does not reportedly have an effect on GFP expression, so it was not removed by the donating laboratory.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, all 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a C57BL/6N genetic background.

Allele

Symbol: Il17a^tm1Bcgen
Name: targeted mutation 1, Biocytogen LLC
MGI accession ID: MGI:5426354
Generation method: Targeted
Attributes: Reporter
Marker symbol: Il17a
Marker name: interleukin 17A
Chromosome: 1
Strain of origin: C57BL/6
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: EGFP is expressed in some naive intraepithelial lymphocytes (IELs) of the small intestine and in activated CD4+ T cells.
Molecular note: An IRES-EGFP-SV40-polyA signal sequence cassette was inserted after the stop codon. A loxP-flanked neomycin cassette is located just after the GFP coding region/3' UTR. It does not have an effect on GFP expression