Mouse dry-eye animal model and application thereof

A mouse model and animal model technology, applied in compound screening/testing, biochemical equipment and methods, preparations for in vivo experiments, etc., can solve long-term, large variability, high-tech conditions of dry eye animal models, etc. problem, to achieve the effect of high repeatability and stable model

Inactive Publication Date: 2020-04-28
QINGDAO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The establishment of these dry eye animal models often requires higher

Method used

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  • Mouse dry-eye animal model and application thereof
  • Mouse dry-eye animal model and application thereof
  • Mouse dry-eye animal model and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Construction of AQP5 - / - Mouse dry eye animal model

[0024] 1. AQP5 - / - Construction and identification of mice

[0025] AQP5 (NCBI ID: 11830) is located on chromosome 15 of the mouse, AQP5 - / - For mouse information and knockout regions see figure 1 A, B, using CRISPR / Cas9 technology to design sgRNA, and knocking out the chr15:99589876-99594444 region with a length of 4569bp gene fragment by applying high-throughput electroporation of fertilized eggs to obtain AQP5 - / - mice. figure 1 C is for AQP5 - / - The sequence identification results of the mouse showed that compared with the wild-type sequence, a total of 4569bp sequence was missing, indicating that the model mouse was successfully established. The mouse has been successfully bred in our institution, and a stable population of homozygous mice has been obtained.

[0026] In the follow-up routine feeding, the PCR amplification method was used to detect AQP5 - / - Knockout effect in mice.

[0027] PC...

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Abstract

The invention provides a stable dry-eye animal model. The dry-eye animal model is constructed through knocking out a gene AQP5 from an animal; and the knockout of the gene AQP5 means knocking out nucleotide fragments of a region 99589876-99594444 of a fifteenth chromosome of a mouse. The dry-eye mouse model provided by the invention is applied to screening or evaluating on effects of dry-eye treatment drugs. The AQP5<-/-> mouse model provided by the invention is high in repeatability and stable in model, researches on dry-eye pathological mechanism and drug intervention are facilitated, and influence on experimental results caused by other influencing factors can be avoided. Furthermore, the AQP5<-/-> mouse model provided by the invention emerges a characteristic change, i.e., reduced tearsecretion after birth, and after birth, lacrimal glands outside orbits emerge a pathologic characteristic, i.e., vacuolar degeneration; and the secretion of tears is free of obvious changes along with increase of ages and is free of sex difference.

Description

technical field [0001] The invention belongs to the technical field of medical animal models, and in particular relates to a method for establishing a dry eye animal model. Background technique [0002] Dry eye refers to a class of diseases that cause tear film instability and / or abnormal ocular surface due to abnormal tear quality, quantity and dynamics, accompanied by ocular symptoms. It is one of the most common diseases in ophthalmology, which can cause ocular surface damage and blurred vision, etc., and affect the quality of life of patients. [0003] Dry eye can be divided into evaporative dry eye, water deficiency dry eye, mucin abnormal dry eye, abnormal tear dynamics dry eye and mixed dry eye. Clinical investigations show that dry eye is a common ocular surface disease, and can cause keratitis, corneal neovascularization, and corneal ulcers, which seriously affect people's quality of life and work efficiency. The common pathophysiological features of dry eye inclu...

Claims

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Application Information

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IPC IPC(8): C12N15/90C12N15/85A01K67/027A61K49/00
CPCC12N15/907C12N15/8509A01K67/0275A61K49/0008A01K2217/075A01K2227/105A01K2267/03
Inventor 陈鹏狄国虎陈豪
Owner QINGDAO UNIV
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