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Construction and application of transgenic human APP double mutant gene Alzheimer's disease mouse model

A technology of APP gene and mouse model, applied in the field of biomedicine, can solve the problems of time lag in pathological and behavioral changes

Inactive Publication Date: 2010-12-15
北京华阜康生物科技股份有限公司
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AI Technical Summary

Problems solved by technology

[0004] At present, a variety of APP mutant transgenic mouse models have been established at home and abroad, such as PDAPP transgenic mouse model, APP23 transgenic mouse model, etc., but most of the models show pathological and behavioral changes later, usually at the age of six months. The above changes only occurred later

Method used

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  • Construction and application of transgenic human APP double mutant gene Alzheimer's disease mouse model
  • Construction and application of transgenic human APP double mutant gene Alzheimer's disease mouse model
  • Construction and application of transgenic human APP double mutant gene Alzheimer's disease mouse model

Examples

Experimental program
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Embodiment 1

[0025] Example 1: Preparation and model identification of APP double mutant transgenic vector and mice:

[0026] 1. Preparation of APP double mutant transgenic vector and mice

[0027] The APP double mutant gene was inserted into the downstream of the PDGF promoter to construct a transgene expression vector (such as figure 1 ). Establishment of APP695 by a simple microinjection procedure V652I / K596N / M597L Transgenic C57BL / 6J mice. Target gene information: Human APP695 V642I / K595N / M596L mutant, corresponding to APP770 V717I / K670N / M671L Build information:

[0028]

[0029] 1. PCR identification of mouse model:

[0030] 1) Cut 0.5-1cm mouse tail and put it in EP tube, add 0.5ml mouse tail lysate, then add 20mg / ml proteinase K 10μl, shake overnight at 55℃;

[0031] 2) The next day, add 300 μl saturated sodium chloride, let stand on ice for 15 minutes, and centrifuge at 13000 rpm for 15 minutes;

[0032] 3) Take 600 μl of the supernatant, add an equal volume of isopropan...

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Abstract

The invention belongs to the field of medicaments and provides construction and application of a transgenic human APP double mutant gene Alzheimer's disease mouse model. The APP double mutant gene Alzheimer's disease mouse model is characterized by having clinical characteristics and pathological changes of human Alzheimer's disease, namely the mouse model expresses A beta (a main component of intracerebral senile plaque) in three months, has A beta accumulation and early senile plaque in neurons in five months, has learning and memory impairment in three months and the like. Compared with other Alzheimer's disease animal models, the model has the characteristics of early illness outburst and obvious clinical and pathological characteristics. Therefore, the construction and application of the APP double mutant gene Alzheimer's disease mouse model provide a tool for researching the pathogenesis of human Alzheimer's disease and developing medicaments and vaccines.

Description

technical field [0001] The invention belongs to the field of biomedicine, and the dementia mouse model of human APP double mutation gene (PDGF-hAPP) is transferred V652I / K596N / M597 ) is an animal model used to study the pathogenesis, drug and vaccine development of Alzheimer's disease (AD). Background technique [0002] Alzheimer's disease (AD) is a common degenerative disease of the central nervous system, the main clinical manifestations are progressive memory loss and cognitive impairment, cognitive dysfunction and slow thinking, and progressive aggravation, It is the most common dementia disease in the elderly population, and its patients account for 50%-70% of all dementia cases. Both the incidence and prevalence of AD increase exponentially with age. According to figures provided by the United Nations, there are more than 50 million elderly people worldwide suffering from AD, and this number is expected to quadruple by 2050. At present, the elderly over 60 years old...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/63A01K67/027A61K49/00
Inventor 杨志伟李万波张连峰方瑾陈炜
Owner 北京华阜康生物科技股份有限公司
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