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PEAR1 gene mutation kit and application thereof

A PEAR1-T4, kit technology, applied in the field of PEAR1 gene mutation kits, to achieve the effect of improving the survival rate

Pending Publication Date: 2021-03-19
PEKING UNIV FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, there is no in vitro model of PEAR1 gene mutation that can simulate the effect of PEAR1 gene mutation on thrombotic diseases in the population.

Method used

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  • PEAR1 gene mutation kit and application thereof
  • PEAR1 gene mutation kit and application thereof
  • PEAR1 gene mutation kit and application thereof

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Experimental program
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Effect test

Embodiment 1

[0036] 1. PEAR1 gene knockout mouse construction strategy and plasmid construction

[0037] Referring to the PEAR1 genome data, it was found that there are multiple transcripts of the PEAR1 gene. The transcripts and protein conserved regions were analyzed, and a TALEN endonuclease that can specifically recognize the DNA of the exon functional region of the PEAR1 gene was designed. Cut PEAR1 genomic DNA, induce DNA damage, and form mutation positions during homologous recombination repair. TALEN plasmids such as figure 1 shown.

[0038] 2. Detection of SSA activity of TALEN

[0039] TALEN plasmid construction, based on the analysis of PEAR1 gene transcripts and protein conservative regions, frameshift mutation design starts before the first conserved region "EMI", so the TALEN target design is on the public exon 2 or 3 at the beginning of ATG.

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Abstract

The invention relates to a PEAR1 gene mutation kit and application thereof. The kit comprises a TALEN endoenzyme or a plasmid for encoding the TALEN endoenzyme. The PEAR1 gene mutation kit disclosed by the invention can be used for successfully mutating a PEAR1 gene, the activity of a PEAR1 protein can be lowered through mutation, and increase of survival rate of pulmonary embolism is facilitatedthrough lowering the activity of the PEAR1 protein.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a PEAR1 gene mutation kit and application thereof. Background technique [0002] Platelet endothelial aggregation receptor-1 (PEAR1) is a transmembrane protein containing 1034 amino acids, mainly expressed on megakaryocytes, platelets and endothelial cells. PEAR1 contains an EMI domain (protein-protein interaction region) outside the cell, 15 repeated sequences similar to epidermal growth factor and multiple tyrosine and proline residues in the cytoplasm. [0003] Among multiple population-based studies, multiple independent studies have found that the PEAR1 gene can regulate platelet function and participate in the occurrence of thrombosis-related events. Several genome-wide association studies (Genome-Wide-Association-Studies, GWAS) found that PEAR1 gene mutations can affect the aggregation ability of platelets. Juraj Sokol et al found that PEAR1 single nucleo...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 崔一民向倩王哲胡琨常国栋
Owner PEKING UNIV FIRST HOSPITAL
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