B6;SJL-Stxbp5^em1Cjln/Mmjax

Stock number: 033363
Product name: Stxbp5(N437S)
Research areas: Cardiovascular Research

Description

Stxbp5(N437S) knock-in mice carry an N437S (asparagine to serine) substitution in the Stxpb5 (syntaxin binding protein 5 or tomosyn) gene. Homozygous mice exhibit decreased plasma von Willebrand factor levels and decreased thrombosis.

Detailed description

Stxbp5 (syntaxin binding protein 5 or tomosyn) gene encodes a protein involved in regulation of platelet granule exocytosis and thrombosis. GWAS (genome-wide association studies) have identified a human SNP in the STXBP5 locus associated with altered thrombosis. The SNP, rs1039084, carries an amino acid substitution, N436S, that is characterized by decreased thrombosis.  Stxbp5(N437S) knock-in mice carry the substitution orthologous to the human N436S variant. Mice homozygous for the mutation are viable and fertile. Homozygous mice exhibit lower plasma von Willebrand factor (vWF) levels, prolonged bleeding time, decreased venous thrombosis and decreased platelet secretion.

Development

CRISPR/cas9 endonuclease-mediated genome editing was used to create an amino acid substitution at position 437 altering asparagine to serine (N437S; AAT to AGC) in exon 13 of the Stxbp5 (syntaxin binding protein 5 (tomosyn)) gene. The mutation is located in the WD40 repeat domain, which is involved in regulation of exocytosis in multiple cell types. Single guide RNA and Cas9 endonuclease were introduced into B6SJLF1-derived fertilized eggs by pronuclear injection. Embryos were transferred to pseudopregnant females, and correctly targeted pups were bred to wild-type mice for germline transmission. The colony was maintained on a mixed C57BL/6 and SJL/J background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Stxbp5^em1Cjln
Name: endonuclease-mediated mutation 1, Charles J Lowenstein
MGI accession ID: MGI:6315230
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Synonyms: Stxbp5(N437S)
Marker symbol: Stxbp5
Marker name: syntaxin binding protein 5 (tomosyn)
Marker synonyms: 0710001E20Rik; 4930565N16Rik; LGL3; LLGL3; mKIAA1006; Nbla04300; tomosyn 1
Chromosome: 10
Strain of origin: (C57BL/6 x SJL)F1
Molecular note: CRISPR/cas9 endonuclease-mediated genome editing was used to create an amino acid substitution at position 437 altering asparagine to serine (N437S; AAT to AGC) in exon 13 of the gene. The mutation is located in the WD40 repeat domain, which is involved in regulation of exocytosis in multiple cell types. The orthologous human variant N436S (SNP rs1039084) is associated with decreased thrombosis.