Construction method and application of spontaneous gastric precancerous lesion animal model

A technology of gastric precancerous lesions and construction methods, applied in the field of animal model construction of spontaneous gastric precancerous lesions, can solve the problems of time-consuming, large individual differences, and low clinical fit

Pending Publication Date: 2022-03-25
GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the complex pathogenesis of gastric precancerous lesions, the existing animal model replication methods are time-consuming, costly, have large individual differences, poor stab...

Method used

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  • Construction method and application of spontaneous gastric precancerous lesion animal model
  • Construction method and application of spontaneous gastric precancerous lesion animal model
  • Construction method and application of spontaneous gastric precancerous lesion animal model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Animal model ATP4a + / - Mouse construction.

[0046] 1.1 Gene editing

[0047] Retrieve the C57BL / 6J mouse gene database MGI (http: / / www.informatics.jax.org / ), and retrieve the wild-type gastric H + -K + - ATPase 4a gene (ATP4a, ATPase, gastric H + / K + exchanging alphapolypeptide), MGI gene number: 88113, NCBI Gene number: 11944, Ensembl: ATP4a (ENSMUSG00000005553). Located in mouse chromosome 7: NC_000073.7, gene start and stop sequence: 30411634~30424959. Its gene structure is as follows figure 1 shown.

[0048] The gene transcript ATP4a-201 (Ensembl number: ENSMUST00000005692.13) has 22 exons, 50 structural domains and features. CRISPR / Cas9 technology knocks out exons exon5-11 (introns between consecutive exons 5-11) in the ATP4a gene transcript of C57BL / 6J mice, such as figure 2 shown.

[0049] After cleavage, connect the cleavage sites 1 and 2 to form the ATP4a silent expression gene after gene knockout, see image 3 . Cas9 mRNA and gRNA were obtained ...

Embodiment 2

[0066] 1. Materials and methods

[0067] 1.1 Animals

[0068] 4-5 weeks healthy SPF grade littermate first generation (F1) wild-type ATP4a + / + and ATP4a + / - Mice, weighing 20-25 g, provided by Shanghai South Model Organism Co., Ltd., experimental animal license number: SCXK 2014-0002, animal qualification certificate number: 312024300003479, self-cultivated to the F3 generation for future use; animal breeding environment: Guangzhou Traditional Chinese Medicine University Animal Experiment Center SPF Animal Laboratory, environmental certification number: SYXK (Guangdong) 2013-0001.

[0069] 1.2 Drugs and reagents

[0070] Anti-rabbit primary antibody MUC2 (lot number: Ab76774), anti-rabbit Ki-67 (lot number: Ab16667), product of abcam USA; AMACR (lot number: NBP1-87168), product of Novous company; anti-mouse p53 (lot number: CBL404), Products of Millipore Company; Immunohistochemistry-related secondary antibodies: Mouse / Rabbit Universal Immunohistochemistry Detection Kit (G...

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Abstract

The invention discloses a construction method and application of a spontaneous gastric precancerous lesion animal model, and relates to the technical field of medical treatment. The gene knockout heterozygous animal ATP4a < +/-> is constructed, and the gene knockout heterozygous animal ATP4a < +/-> is used as the gastric precancerous lesion animal model, can spontaneously generate gastric mucosa full-thickness tissue heterotypic incomplete intestinal epithelial metaplasia lesion, can be used as the gastric precancerous lesion animal model, and can be used as a gastric precancerous lesion animal model. The compound is suitable for research and development of gastric precancerous lesion pathogenesis and efficacy and mechanism of new drugs for resisting chronic atrophic gastritis.

Description

technical field [0001] The invention relates to the technical field of medical treatment, in particular to a method for constructing an animal model of spontaneous gastric precancerous lesions and its application. Background technique [0002] Gastric cancer is a malignant tumor originating from the gastric mucosal epithelium. Due to changes in diet structure, increased work pressure, and Helicobacter pylori infection, gastric cancer tends to be younger. Gastric cancer can occur in any part of the stomach, and more than half of them occur in the gastric antrum. The greater curvature, the lesser curvature, and the anterior and posterior walls can be involved. The vast majority of gastric cancer is adenocarcinoma, with no obvious symptoms in the early stage, or non-specific symptoms such as epigastric discomfort and belching, which are often similar to the symptoms of chronic gastric diseases such as gastritis and gastric ulcer, and are easily overlooked. [0003] Precancero...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/12A01K67/027
CPCC12N15/8509C07K14/47A01K67/0276A01K2217/075A01K2227/105A01K2267/0331A01K2267/0368
Inventor 潘华峰倪家慧刘伟赵自明刘远亮杨良俊张丹田雯李嘉丽
Owner GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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