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PirB gene-knock-in mouse animal model and construction method thereof

A gene knock-in and animal model technology, applied in the fields of genetics and biology, can solve problems such as low transfection efficiency and PirB functional limitations

Active Publication Date: 2020-02-11
XIAN MEDICAL UNIV
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Problems solved by technology

The former is affected by the ability to penetrate the blood-brain barrier and the efficiency of inhibitors, while the latter has a relatively low transfection efficiency in primary cells and in vivo transfection, which greatly limits the study of the function of PirB

Method used

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  • PirB gene-knock-in mouse animal model and construction method thereof
  • PirB gene-knock-in mouse animal model and construction method thereof
  • PirB gene-knock-in mouse animal model and construction method thereof

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Embodiment Construction

[0026] The invention will be further elaborated below in conjunction with the accompanying drawings and specific embodiments.

[0027] The present invention constructs a PirB gene knock-in mouse animal model, and knocks the PirB gene into intron 1 of the ROSA26 gene of C57BL / 6J mice. Specifically, a CAG promoter-loxP-Stop-loxP-Kozak-mouse PirB CDS-polyA gene cassette was constructed and cloned into intron 1 of the ROSA26 gene.

[0028] The ROSA26 gene (NCBI Reference Sequence: NR_011095.2) is located on chromosome 7 of the mouse.

[0029] The construction method of the mouse animal model that PirB gene knocks in of the present invention, specifically implement according to the following steps:

[0030] Step 1. According to the mouse ROSA26 gene (Gene Bank: NR_027008.1)) sequence, use Cas-Desigher software to design the gRNA target sequence in intron 1, and search the mouse DBM-DB genome database gene, and use CRISPR off-target effect The software Cas-OFFinder detects potenti...

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Abstract

The invention discloses a PirB gene-knock-in mouse animal model, which comprises gRNA1 and gRNA2 for determining specific target sites to be knocked in by PirB gene, wherein the PirB gene is knocked into an intron 1 of ROSA26 gene of a C57BL / 6J mouse; the gene sequence of the gRNA1 is shown in SEQ ID NO.1; and the gene sequence of the gRNA2 is shown in SEQ ID NO.2. The invention further specifically discloses a construction method of the mouse animal model, the mouse animal model provides a good foundation for research on PirB gene function and in-vivo verification. Hybridization of PirB-knock-in animal model with different types of Cre mice can be used to study the functions of PirB in different organs or different cell types or different disease models.

Description

Technical field: [0001] The invention belongs to the field of genetics and biotechnology, and specifically relates to a PirB gene knock-in mouse animal model, and also relates to a method for constructing the mouse animal model. Background technique [0002] Mouse-derived paired immunoglobulin-like receptor B (PirB), which is the homologous gene of human immunoglobulin-like receptor B2 (human leukocyte immunoglobulin (Ig)-like receptor B2, LILRB2), PirB The gene (NCBI reference sequence: NM_011095.2) is located at the proximal end of chromosome 7 of the mouse, with a total size of 8.97kb, ​​encoding 841 amino acids, and 15 exons have been identified so far. The start code of exon 1 is ATG, The stop codon for exon 15 is TGA (transcript: Pirb-201ENSMUST00000078451.6). PirB protein belongs to type I transmembrane glycoprotein, which contains an extracellular segment composed of six immunoglobulin-like domains (domain, D) (D1-D6), a hydrophobic transmembrane segment, and three ...

Claims

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Application Information

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IPC IPC(8): A01K67/027C12N15/90C12N15/85C12N15/12
CPCA01K67/0275A01K2217/072A01K2227/105A01K2267/03A01K2267/0387C07K14/70503C12N15/8509C12N15/907C12N2800/107C12N2800/30
Inventor 张晓华苟兴春王晓龙姜朋涛程江红邱忠营
Owner XIAN MEDICAL UNIV
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