NOX2 gene defect rd1 mouse model as well as preparation method and application thereof

A mouse model, gene defect technology, applied in pharmaceutical formulations, preparations for in vivo experiments, compound screening/testing, etc., can solve the problems of inoperability of apocynin retinal degeneration, inaccurate action mechanism of apocynin, and poor NOX2 inhibitory effect. , to achieve the effect of delaying the loss of photoreceptor cells in the outer nuclear layer of the retina, clear screening and development, and clear mechanism of action

Pending Publication Date: 2022-05-27
BEIJING TONGREN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV +1
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Problems solved by technology

Document "Sorce, S., K.H.Krause, and V.Jaquet.Targeting NOX enzymes in the centralnervous system: therapeutic opportunities. Cell Mol Life Sci.2012; 69(14):2387-407." It is also mentioned that apocynin has The inaccurate mechanism of action, lack of specificity and poor inhibitory effect on NOX2, and the high failure rate of the previous use of antioxidant drugs in the treatment of hereditary retinal degeneration further prove that apocynin is not feasible in the treatment of retinal degeneration

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  • NOX2 gene defect rd1 mouse model as well as preparation method and application thereof
  • NOX2 gene defect rd1 mouse model as well as preparation method and application thereof
  • NOX2 gene defect rd1 mouse model as well as preparation method and application thereof

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Embodiment 1

[0036] Example 1: NOX2 gene-deficient rd1 mouse model and its preparation method

[0037] 1. NOX2 knockout (NOX2KO) mice were crossed with rd1 mice to obtain F1 generation mice

[0038] Four male NOX2 knockout mice with genotype gp91phox- / Y rd1W / W (that is, B6.129S-Cybb with NOX2 knockout on the X chromosome) tm1Din / J mice (purchased from Jackson Laboratory, JAX code: 002365) and eight female rd1 mice (purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.) with genotype gp91phox+ / +rd1+ / +, obtained Male cross-bred F1 mice with genotype gp91phox+ / Y rd1W / + and female cross-bred F1 mice with genotype gp91phox+ / -rd1w+.

[0039] In order to verify the correctness of the genotypes of the F1 generation mice, the genomes of the F1 generation mice were amplified by PCR using the primers in Table 1, and the amplified products were analyzed by gel electrophoresis. The analysis results are shown in figure 2 .

[0040] Depend on figure 2 It can be seen that the ...

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Abstract

The invention relates to an NOX2 gene defect rd1 mouse model as well as a preparation method and application thereof, and belongs to the technical field of biology. The invention provides a method for preparing an NOX2 gene defect rd1 mouse model, which comprises the following steps: hybridizing and screening an NOX2 gene knockout mouse and an rd1 mouse to obtain the NOX2 gene defect rd1 mouse model. The loss of retinal outer nuclear layer photoreceptor cells of the NOX2 gene defect rd1 mouse model prepared by using the method is obviously delayed, the activation of microglia cells is obviously inhibited, and the expression of NOX2 in the microglia cells is obviously reduced, so that the NOX2 activation in the microglia cells is further verified to be a pathogenic mechanism of hereditary retinal degeneration; in addition, the invention provides a basis for screening and researching and developing medicines for treating and / or preventing retinal degeneration, which are clearer in action mechanism, stronger in specificity and better in market and clinical application prospect, and has a great application prospect.

Description

technical field [0001] The invention relates to a NOX2 gene-deficient rd1 mouse model, a preparation method and application thereof, and belongs to the field of biotechnology. Background technique [0002] Primary retinitis pigmentosa (RP) is a group of hereditary eye diseases that cause progressive photoreceptor cell apoptosis. Although gene and stem cell therapy has made some progress in the treatment of primary retinitis pigmentosa, due to the complexity of the RP phenotype or the potential problems of the technology itself, its widespread clinical application is not realistic. It is still very important to study the common pathological mechanisms that occur downstream of gene variation. Even if the pathogenic cause (gene mutation) of the disease cannot be fundamentally changed, drug therapy targeting the core link of the common pathogenic mechanism can still delay the loss of photoreceptor cells, delay the blinding time of patients, and improve their quality of life. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027A61K49/00
CPCA01K67/027A61K49/0008A01K2217/075A01K2227/105A01K2267/03
Inventor 曾惠阳刘谦周健武珅
Owner BEIJING TONGREN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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