B-hSAS1B mice_Biocytogen Pharmaceuticals (Beijing) Co., Ltd._Biocytogen, drug development, antibody discovery

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동물/세포 모델

동물/세포모델

Humanized Mouse Models

Other Humanized Mice

Text

B-hSAS1B mice

Strain Name	C57BL/6N-Astl^{tm1(ASTL)Bcgen}/Bcgen	Common Name	B-hSAS1B mice
Background	C57BL/6N	Catalog number	112722
Related Genes	OOMD11, OZEMA11, SAS1B
NCBI Gene ID	431705

Description

Ovastacin (ASTL, astacin-like metallo-endopeptidase and Oocyte astacin) also known as SAS1B is a 50 kDa member of the peptidase M12A family of proteins，which is comprised of a signal peptide at the N-terminus, pro-peptide, proteinase domain containing a Hex-box catalytic site, and a unique C-terminal domain.
Ovastacin is an oocyte-specific oolemmal receptor involved in sperm and egg adhesion and fertilization, and it plays a role in the polyspermy inhibition.
The exons 6-9 of mouse Sas1b gene that encode extracellular domain are replaced by human counterparts in B-hSAS1B mice. The genomic region of mouse Sas1b gene that encodes signal peptide, transmembrane domain and cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric SAS1B expression is driven by endogenous mouse Sas1b promoter, while mouse Sas1b gene transcription and translation will be disrupted.
SAS1B was detectable in kidney, lung(weak expression) and liver of wild-type C57BL/6N mice and homozygous B-hSAS1B mice, as the antibody can cross-recognize both human and mouse SAS1B.
Mouse Sas1b mRNA was only detectable in wild-type C57BL/6N mice. Human SAS1B mRNA was only detectable in homozygous B-hSAS1B mice.
Application: This product is used for pharmacodynamics and safety evaluation of drugs.

Targeting strategy

Gene targeting strategy for B-hSAS1B mice. The exons 6-9 of mouse Sas1b gene that encode extracellular domain are replaced by human counterparts in B-hSAS1B mice. The genomic region of mouse Sas1b gene that encodes signal peptide, transmembrane domain and cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric SAS1B expression is driven by endogenous mouse Sas1b promoter, while mouse Sas1b gene transcription and translation will be disrupted.

mRNA expression analysis in humanized B-hSAS1B mice

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Species specific analysis of SAS1B gene expression in wild-type C57BL/6N mice and homozygous humanized B-hSAS1B mice by RT-PCR. Ovary was collected from wild-type C57BL/6N mice (+/+) and homozygous B-hSAS1B mice (H/H). Mouse Sas1b mRNA was detectable only in wild-type C57BL/6N mice. Human SAS1B mRNA was detectable only in homozygous B-hSAS1B mice, but not in wild-type C57BL/6N mice.

Protein expression analysis

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Strain specific SAS1B expression analysis in homozygous B-hSAS1B mice by western blot. Heart, liver, lung and kidney were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hSAS1B mice (H/H), and analyzed by western blot with anti-SAS1B antibody(R&D，AF8549). m/hSAS1B was detectable in kidney, lung(weak expression) and liver of wild-type C57BL/6N mice (+/+) and homozygous B-hSAS1B mice.