B6;C-Stxbp1^{tm1.1(STXBP1)Bpro}/Mmjax

Stock number: 038753
Product name: STXBP1 hu

Research areas: Mouse/Human Gene Homologs, Neurobiology Research

Description

Humanized STXBP1 mice express a knock-in/knock-out mutation that results in replacement of the murine Stxbp1 (syntaxin binding protein 1) sequence, from upstream of exon 1 to the end of exon 19, with human sequence from upstream of exon 1 to the final exon. These mice may be useful for delivering and evaluating the efficacy of biological therapeutics, specifically for neurological diseases.

Detailed description

Stxbp1 (syntaxin binding protein 1) encodes a protein involved in the release of neurotransmitters through the binding of syntaxin. Mutations in Stxbp1 are associated with epilepsy, intellectual disability and movement disorders. STXBP1 hu knock-in/ knock-out mice express a BAC that replaces the Stxbp1 (syntaxin binding protein 1) murine sequence upstream of exon 1 to the end of exon 19 with the human sequence from upstream of exon 1 to the final exon. Mice homozygous for the mutation are viable and fertile, however, viability is reduced in homozygote males. RT-qPCR confirms a 50% reduction of murine Stxbp1 RNA in heterozygous mice and the expression of human STXBP1 RNA. These mice may be useful for delivering and evaluating the efficacy of biological therapeutics, specifically for neurological diseases.

Development

The humanized STXBP1 knock-in/knock-out targeting vector contains from 5' to 3' a loxP site, an FRT-flanked neomycin cassette, a human genomic DNA BAC encoding STXBP1 gene, an F3-flanked hygromycin, followed by a second loxP site used to replace the endogenous murine sequence from approximately 5.4kb upstream of exon 1 through to near to the end of exon 19 with the human genomic region from approximately 4.0kb upstream of exon 1 (Transcript ENST00000373299.5) through to the end of the final exon of human transcript (ENST00000636962.2). This humanized region may also express the short 35 residue isoform of PTRH1 (Uniprot: A0A286YER0) on chromosome 2. The insertion results in a knock-out of Stxbp1.The construct was electroporated into C57BL/6-derived embryonic stem (ES) cells (Ozgene). Correctly targeted ES cells were injected into recipient blastocysts. Chimeric mice were bred to Flp deleter mice (Ozgene) on the C57BL/6 background to remove the neomycin and hygromycin cassettes. The FLP allele was later bred out of the colony. To improve breeding, mice were bred to BALB/c mice; the strain is maintained on the mixed background. Upon arrival at The Jackson Laboratory, 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: Stxbp1^{tm1.1(STXBP1)Bpro}
Name: targeted mutation 1.1, Benjamin Prosser
MGI accession ID: MGI:7524737
Generation method: Targeted
Attributes: Null/Knockout; Inserted expressed sequence; Humanized sequence
Synonyms: STXBP1 hu
Marker symbol: Stxbp1
Marker name: syntaxin binding protein 1
Marker synonyms: ANC18HA; DEE4; MUNC18-1; Munc-18a; nsec1; n-sec1; NSEC1A; P67; rbSec1; Rb-sec1; rbSec1A; rbSec1B; Sec1; Sxtbp1; UNC18; unc18-1; unc-18A; Unc18h
Chromosome: 2
Strain of origin: C57BL/6
Molecular note: The targeting vector contains from 5' to 3' a loxP site, an FRT-flanked neomycin cassette, a human genomic DNA BAC encoding STXBP1 gene, an F3-flanked hygromycin, followed by a second loxP site used to replace the endogenous murine sequence from approximately 5.4kb upstream of exon 1 through to near to the end of exon 19 with the human genomic region from approximately 4.0kb upstream of exon 1 (Transcript ENST00000373299.5) through to the end of the final exon of human transcript ENST00000636962.2. This humanized region may also express the short 35 residue isoform of PTRH1 (Uniprot: A0A286YER0) on chromosome 2. The insertion results in a knock-out of Stxbp1. Flp-mediated recombination removed the neo and hygro cassettes.