NOD.Cg-Myd88^tm1Hlz H2^b/JkraJ

Stock number: 039018
Product name: NOD.B10LSL-/-
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

The NOD.B10MyD88^LSL knock-in/knock-out allele, backcrossed onto a NOD.B10 genetic background (Stock No. 002591), is a functional null prior to Cre recombinase exposure. This congenic strain has the MHC H2^b haplotype from the C57BL/10SnJ strain with mice exhibiting some but not all of the immune dysfunctions of NOD/LtJ (Stock No. 001976) mice. These mice allow Cre recombinase-inducible restoration of functional MYD88 expression and may be useful for studying cell-type specific function of MYD88 in various inflammatory and autoimmune diseases.

Note, the same MyD88^LSL allele is available on a congenic C57BL/6 background as Stock No. 028939.

Detailed description

Myeloid differentiation primary response gene 88 (MyD88) codes for a cytosolic adapter protein that is essential in signal transduction of the toll-like receptor (TLR) signaling pathway involved in innate and adaptive immune responses. Myd88 mutations have been linked to autoimmune diseases such as lupus, multiple sclerosis (MS) and Sjögren's syndrome (SS).

These NOD.B10Myd88^LSL mice carry an knock-in/knock-out (KIKO) allele that has a floxed transcriptional termination element (loxP::En2 splice-acceptor::3xSTOP::IRES::lacZ-pA::DSE::loxP-NeoR-loxP), that prevents expression of Myd88, on a NOD.B10 (Stock No.002591) genetic background. The NOD.B10 (or NOD.H2-b) strain has the MHC H2^b haplotype from the C57BL/10SnJ strain with mice exhibiting some but not all of the immune dysfunctions of NOD/LtJ (Stock No.001976) mice. Female mice of this strain do not develop diabetes insulitis, cyclophosphamide-induced diabetes or spontaneous diabetes, but do develop lymphocytic infiltrates in the kidneys, lungs, lacrimal glands, and the submandibular glands as well as other SS pathologies ~26 weeks of age. Following cre-mediated recombination the entire floxed region is deleted allowing for recovery of MYD88 expression and function.

Homozygous NOD.B10Myd88^LSL mice are viable and fertile. NOD.B10Myd88^LSL mice may be useful for studying cell-type specific function of MYD88 in autoimmune and inflammatory diseases.

Note, the same Myd88^LSL allele is available on a congenic C57BL/6 background as Stock No.028939.

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. It should be noted that the phenotype of these NOD.B10 congenic mice could vary from that originally described on a C57BL/6 genetic background. We may modify the strain description if necessary, as published results become available.

Development

The NOD.B10Myd88^LSL mouse model was generated by backcrossing the C57BL6-congenic Myd88^LSL allele (Stock No. 028939) onto the NOD.B10 background (Stock No. 002591).

The NOD.B10, or NOD.H2-b, congenic mouse line was made by transferring the H2b haplotype from C57BL/10SnJ (Stock No. 000666) mice to the NOD/Lt (Stock No. 001976) strain.

The MyD88^LSL allele was created in the laboratory of Dr. Bernhard Holzmann (Technische Universität München) by inserting a floxed transcriptional termination element between exons 1 and 2 of the myeloid differentiation primary response gene 88 locus (Myd88) on chromosome 9. The targeting vector contained (from 5' to 3') a loxP site, an engrailed-2 splice acceptor site followed by stop codons in all reading frames (En-2/stp), an internal ribosomal entry site (IRES), a β-galactosidase sequence followed by a polyadenylation signal (lacZ-pA), a downstream element (DSE) and a loxP-flanked neomycin resistance cassette. The construct was electroporated into 129P2/OlaHsd-derived E14.1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts, and chimeric mice were bred with C57BL/6N mice for germline transmission and to establish the MyD88^LSL mice. The donating investigator reported that the MyD88^LSL colony was backcrossed to C57BL/6N for a total of more than ten generations prior to sending heterozygous males to The Jackson Laboratory Repository in 2016. The C57BL/6-congenicMyd88^LSL colony is Stock No. 028939.

Dr. Jill M. Kramer acquired some Stock No. 028939 animals, on the C57BL/6 congenic background, and then backcrossed them to NOD.B10 mice (Stock No. 002591) for at least 5 generations using The Jackson Laboratory speed congenic protocol. The resulting NOD.B10-congenic strain is called NOD.Cg-Myd88^tm1Hlz H2b/JkraJ, Stock No. 039018. To establish our live colony, an aliquot of frozen sperm was used to fertilize NOD/ShiLtJ (Stock No. 001976) oocytes.

In 2024, a 48 SNP (single nucleotide polymorphism) panel analysis, with 43 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. One of the markers on Chromosome 9 (~101 Mb) was found to be segregating in some MyD88^LSL heterozygotes, possibly due its proximity to the Myd88 locus (~119 Mb).

Allele

Symbol: H2^b
Name: b variant
MGI accession ID: MGI:3579319
Generation method: Not Applicable
Marker symbol: H2
Marker name: histocompatibility-2, MHC
Marker synonyms: H-2; MHC-II
Chromosome: 17
Strain of origin: Not Applicable
General note: The b variant has been observed in the following strains: C57BL/10, C57BL/10SnJ, BXSB/MpJ, C57BL/6J, C57L/J, LP/J, 129P3/J, 129P1/ReJ.

Allele

Symbol: Myd88^tm1Hlz
Name: targeted mutation 1, Bernhard Holzmann
MGI accession ID: MGI:5442766
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Null/Knockout
Synonyms: Myd88^LSL; MyD88^OFF
Marker symbol: Myd88
Marker name: myeloid differentiation primary response gene 88
Marker synonyms: IMD68; MYD88D; WM1
Chromosome: 9
Strain of origin: 129P2/OlaHsd
Molecular note: An intron gene trap cassette, containing a loxP site, Engailed 2 splice acceptor/STOP (En-2/stp), IRES, lacZ, downstream element (DSE) and floxed neo cassette, was targeted to intron 1. This allele is both a knock-out and conditional-ready allele, as it can be reverted back to wild-type expression through cre-mediated recombination. Western blot analysis confirmed the absence of protein expression in bone marrow derived macrophage stimulated with (S)-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-N-palmitoyl-(R)-Cys-(S)-Ser-(S)-Lys4-OH (P3C).