Construction method for constructing Alzheimer's disease animal model and nucleic acid composition and application thereof

A technology for Alzheimer's disease and nucleic acid composition, applied in the field of genetic engineering, which can solve the problems of limiting the use of models and not conforming to the clinical disease progression process of AD patients

Pending Publication Date: 2022-01-21
GEMPHARMATECH CO LTD
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  • Application Information

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Problems solved by technology

However, the symptoms of 3xTg mice progressed to cognitive impairment first and then pathological changes in the brain, which did not conform to the clinical disease progression process of AD patients
In addition, since the existing mature AD models are all donated to the Jackson Laboratory by scientific research institutions, they need to obtain the permission of the donor and only sell them to non-profit institutions (universities and other scientific research institutes), which limits the ability of researchers, especially AD. The use of models by pharmaceutical R&D companies

Method used

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  • Construction method for constructing Alzheimer's disease animal model and nucleic acid composition and application thereof
  • Construction method for constructing Alzheimer's disease animal model and nucleic acid composition and application thereof
  • Construction method for constructing Alzheimer's disease animal model and nucleic acid composition and application thereof

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Experimental program
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Effect test

Embodiment 1

[0057] A method for constructing a mouse model of Alzheimer's disease includes: linking hAPP 5'UTR and mutant hAPP cDNA (carrying Swedish and Indiana mutations) with the Thy1.2 promoter (Thy1 exon2) to form a targeting vector 1. Link the human PSEN1 cDNA carrying the M146L and L286V mutations with the Thy1.2 promoter to form the targeting vector 2. Then, the targeting vector 1 and the targeting vector 2 were simultaneously injected into C57BL / 6J embryos to obtain FAD 4T mouse model. Specific steps are as follows.

[0058] 1. Construction of targeting vectors Thy1-APP and Thy1-PSEN1.

[0059] The hAPP cDNA (including APP 5'UTR) carrying the Swedish mutation and the Indiana (V642F) mutation and the hPSEN1 cDNA carrying the M146L and L286V mutations were respectively cloned into Exon2 (exon 2) downstream of the Thy1 promoter to obtain the targeting vector Thy1 -APP (targeting vector 1) and Thy1-PSEN1 (targeting vector 2), see image 3 .

[0060] 1.1 Preparation of targeting ...

Embodiment 2

[0103] 2. Pathological detection of Alzheimer's disease mouse model.

[0104] 2.1FAD 4T Detection of Aβ plaques in brain regions of mice (same as in Example 1).

[0105] Take FAD of different ages 4T The brains of mice and WT mice were dehydrated, embedded, and sectioned, and immunohistochemical staining was performed to detect the expression of Aβ plaques in the mouse brain regions. The result is as Figure 12 , Figure 13 Shown: FAD at 1.5 weeks of age 4T Aβ plaques can be detected in the cortex and hippocampus of the mouse brain, and as the age increases, the Aβ plaques gradually increase, that is, the increase of Aβ deposition is age-dependent. Aβ plaques increased from the initial cortex and hippocampus to the thalamus and olfactory bulb regions, and a small amount of Aβ plaques appeared in the midbrain.

[0106] 2.2FAD 4T Mouse glial cell status detection.

[0107] FAD 4T The brains of mice and WT mice were dehydrated, embedded, and sectioned, and immunofluoresc...

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Abstract

The invention discloses a construction method for constructing an Alzheimer's disease animal model and a nucleic acid composition and application thereof, and relates to the technical field of genetic engineering. According to the invention, Thy1-hAPP Sweep, Indiana and Thy1-hPSEN1 M146V and L286V are inserted into a target animal genome and are specifically expressed in a brain region, so that an animal model which can be stably inherited and has a spontaneous classic phenotype of Alzheimer's disease (AD) is obtained. Amyloid protein deposition occurs when the animal model is 1.5 months old, and the amyloid protein deposition increases along with age and is age-dependent. Pathologically along with glial cell hyperplasia, the appearance time is earlier than that of a common AD model. Besides, on the basis that the animal model generates a large amount of A beta deposition, the cognitive and memory ability declines when the animal model is 5 months old, and a convenient and effective way is provided for function analysis of AD related genes, research of disease pathogenesis and drug research and development.

Description

technical field [0001] The present invention relates to the technical field of genetic engineering, in particular to a construction method for constructing an animal model of Alzheimer's disease and its nucleic acid composition and application. Background technique [0002] Alzheimer's disease (AD), also known as senile dementia, is a degenerative disease of the central nervous system. The main manifestations of the disease are neuropsychiatric symptoms such as progressive cognitive impairment, memory impairment, and language impairment, which seriously affect the quality of life of patients. According to the "World Alzheimer's Disease Annual Report 2018" released by the International Alzheimer's Association, there were more than 50 million AD patients worldwide in 2018, and it is expected to reach 74.7 million by 2030 and increase to 1.315 by 2050. billion people. At present, it is generally believed that the deposition of amyloid plaques outside the nerve cells and the f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027
CPCC12N15/8509A01K67/0278A01K2267/0312A01K2227/105
Inventor 琚存祥
Owner GEMPHARMATECH CO LTD
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