B6(Cg)-Siglece^{tm1.1(SIGLEC16)Avrk}/J

Stock number: 032009
Product name: E16
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

These E16 floxed knock-in mice express human SIGLEC16 activating receptor domain from the endogenous mouse Siglece locus. They are suitable for use in applications related to the study of paired Siglec receptors.

Detailed description

The targeted Siglece gene encodes a putative adhesion molecule that mediates sialic-acid dependent binding to cells, and is expressed on innate immune cells. These E16 mice carry a mutation of Siglece in which exon 4 of the endogenous mouse gene has been replaced with cDNA encoding the transmembrane and cytosolic domains of human SIGLEC16 gene, and loxP sites flanking exons 4 and 5. The human SIGLEC16 activating receptor domain replaced the mouse inhibitory domain encoded by exon 4 of the mouse Siglece gene. Expression of human SIGLEC16 is detected in blood neutrophils and macrophages from the liver and spleen of mutant mice. In blood neutrophils from homozygotes, analyzed by flow cytometry, human SIGLEC16 expression levels are similar to the endogenous mouse protein levels. Bacterial survival (E. coli) in blood from homozygotes is significantly reduced when compared to survival in blood from wildtype mice. Macrophages from homozygotes produce increased levels of IL6 and decreased levels of IL10 compared to wildtype controls. Mice that are homozygous for the targeted mutation are viable and fertile.

Development

A targeting vector containing cDNA encoding the transmembrane and cytosolic domains of human SIGLEC16 gene followed by human growth hormone polyadenylation sequence and a FRT-site flanked neo cassette with an upstream loxP site was designed to insert into exon 4 of the endogenous mouse Siglece gene. A second loxP site was inserted downstream of exon 5. The construct was electroporated into unspecified C57BL/6 derived embryonic stem (ES) cells (GenOway). Correctly targeted ES cells were injected into albino B6(Cg)-Tyr^c-2J/J blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were crossed to unspecified mice expressing FLP recombinase on the C57BL/6 background (GenOway) to excise the FRT-site flanked neo cassette. The mice were then backcrossed to C57BL/6 for 10 generations. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6 genetic background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Siglece^{tm1.1(SIGLEC16)Avrk}
Name: targeted mutation 1.1, Ajit Varki
MGI accession ID: MGI:6195824
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Inserted expressed sequence; Humanized sequence
Synonyms: E16; Siglec-E16
Marker symbol: Siglece
Marker name: sialic acid binding Ig-like lectin E
Marker synonyms: AIRM1; AIRM-1; CD328; CD329; CDw328; CDw329; D-siglec; FOAP-9; mSiglec-E; OBBP-LIKE; p75; p75/AIRM1; QA79; RGD1563073; S2V; SAF2; SIGLEC19P; Siglec5; SIGLEC-7; Siglec8; SIGLEC-8; SIGLEC8L; siglec-9; SIGLECL1; SIGLECP2; Siglec-XII; SLG
Chromosome: 7
Strain of origin: C57BL/6
Molecular note: The targeting vector contains from 5' to 3' a loxP site, cDNA encoding the transmembrane and cytosolic domains of human SIGLEC16 gene, a human growth hormone polyadenylation sequence and an FRT-site flanked neo cassette inserted into exon 4. A second loxP site is inserted downstream of exon 5. Flp-mediated recombination removed the FRT-flanked neo cassette. The human SIGLEC16 activating receptor domain replaces the mouse inhibitory domain encoded by exon 4 of the mouse gene. Expression of human SIGLEC16 is detected in blood neutrophils and macrophages from the liver and spleen of mutant mice.