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Mouse model with PIG-A gene specific knockout as well as construction method and application of mouse model

A mouse model, construction method technology, applied in other directions of inserting foreign genetic material, application, genetic engineering, etc., can solve the problem of typical clinical manifestations such as no observed increase in the proportion of PNH

Pending Publication Date: 2022-02-18
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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Problems solved by technology

However, no increase in the proportion of PNH and typical clinical manifestations such as anemia, hemoglobinuria, and thrombus have been observed in either the mouse model or the macaque model, so it cannot be really used in PNH-related research

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  • Mouse model with PIG-A gene specific knockout as well as construction method and application of mouse model
  • Mouse model with PIG-A gene specific knockout as well as construction method and application of mouse model
  • Mouse model with PIG-A gene specific knockout as well as construction method and application of mouse model

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Embodiment Construction

[0035] Unless otherwise defined, the technical terms used in the following embodiments have the same meaning as commonly understood by those skilled in the art to which the present invention belongs. The test reagents used in the following examples, unless otherwise specified, are conventional biochemical reagents; the experimental methods, unless otherwise specified, are conventional methods.

[0036] The present invention is described in detail below in conjunction with embodiment.

[0037] 1. Construction method of mouse model with specific knockout of Pig-a gene

[0038] 1.1 Construction of Pig-a gene-specific knockout vector

[0039] Insert LoxP in the same direction on both sides of exon 3 and exon 5 of the Pig-a gene, construct the targeting vector and verify it, and then carry out plasmid extraction and linearization. For the targeting scheme, see figure 1 . It should be noted that the linearization processing technology is a common technology in this field, and the...

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Abstract

The invention provides a mouse model with PIG-A gene specific knockout as well as a construction method and application of the mouse model. The mouse model with PIG-A gene specific knockout of a hematopoietic system is constructed by using Vav-iCre. The detection of various indexes of mice proves that the model not only has stable GPI deficiency, but also has disease phenotypes of intravascular hemolysis, is an ideal PNH animal model, and can be used in related researches of PNH pathogenesis and treatment.

Description

technical field [0001] The invention belongs to the technical field of animal experiment models and their preparation, and in particular relates to a mouse model with specific knockout of PIG-A gene and its construction method and application. Background technique [0002] Paroxysmal nocturnal hemoglobinuria (PNH) is a disease caused by hematopoietic stem cell membrane defects caused by acquired somatic mutations. The molecular mechanism of PNH is the mutation of the phosphatidylinositol glycan-A (Pig-a) gene on the X chromosome of hematopoietic stem cells, which leads to the synthesis obstacle of glycosylphosphatidylinositol (GPI), which makes the GPI anchor protein ( GPI-AP) deficiency, the main clinical manifestations are hemolytic anemia, thrombosis tendency and bone marrow failure. [0003] At present, it is believed that a simple PIG-A gene mutation is not enough to cause PNH abnormal clones to gain a proliferative advantage and lead to the onset of PNH. The pig-a kno...

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Application Information

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IPC IPC(8): C12N15/89C12N15/85C12N15/12A01K67/027A61K49/00
CPCC12N15/8509C12N15/89C07K14/47A01K67/0276A61K49/0008C12N2800/107A01K2217/075A01K2227/105A01K2267/0381
Inventor 付蓉刘惠陈颖莹李丽燕王红蕾
Owner GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV