B6.129S4-Il17a^tm1.1Lky/J

Stock number: 032406
Product name: il17a^Smart; Smart-17A
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

il17a^Smart/SMART-17A (Surface MARker for Transcription-17A) mice express a human low-affinity human nerve growth factor receptor (hNGFR) marker on the surface of cells expressing mouse Il17a. Endogenous IL17A cytokine production is retained, and phenotype is unaltered aside from the expression of hNGFR. This strain enables the detection and isolation of intact and viable IL17A-producing cells in high purity.

Detailed description

il17a^Smart/SMART-17A (Surface MARker for Transcription-17A) mice express a human low-affinity human nerve growth factor receptor (hNGFR) marker on the surface of cells expressing mouse Il17a. The bi-cistronic messenger RNA reporter approach retains the endogenous IL17A cytokine production and leaves the reporter mouse line phenotypically unaltered except for the expression of the hNGFR marker.

This mouse strain enables the detection and isolation of intact and viable IL17A producing cells in high purity. Tagged cells are then available for studies of infectious and autoimmune disease models in which the production of IL17A has been linked to various outcomes.

Efficacy of the allele was verified by demonstrating that hNGFR expression is specifically induced in CD4⁺ cells in vitro only under Th17 polarizing conditions and that intracellular IL17A is found almost entirely within the hNGFR⁺ population. The hNGFR reporter accurately marks 98% of Th17 cells identified using standard methods of in vitro re-stimulation and intracellular cytokine staining.

Development

A 3.1kb selection/reporter cassette composed of a reversed loxP-neomycin-loxP segment, IRES, low-affinity human nerve growth factor receptor (p75 LNGFR; CD271), and SV40 poly-adenylation signal (pA) was introduced to the 3'UTR of the mouse Il17a gene. The targeting vector was introduced to PC3 129S4/SvJae-Tg(Prm-cre)70Og-derived embryonic stem (ES) cells expressing Cre recombinase from the protamine promoter. The neomycin cassette was deleted in the male germline by breeding male chimeras to C57BL/6J females. Mice carrying the il17a^Smart/Smart-17A allele were backcrossed to C57BL/6J for ten generations by the donating laboratory.

Allele

Symbol: Il17a^tm1.1Lky
Name: targeted mutation 1.1, Richard M Locksley
MGI accession ID: MGI:5439016
Generation method: Targeted
Synonyms: Smart-17A
Marker symbol: Il17a
Marker name: interleukin 17A
Marker synonyms: Ctla8; Ctla-8; Il17; IL-17; IL-17A; ILA17
Chromosome: 1
Strain of origin: 129S4/SvJae-Tg(Prm-cre)70Og
Molecular note: The 3' untranslated region (UTR) was modified to include a floxed neo cassette, an internal ribosomal entry site (IRES) and a non-signaling form of the human nerve growth factor (NGFR) gene. This results in IRES mediated translation of NGFR when the endogenous locus is activated. Cre-mediated recombination removed the neo cassette.