Method for establishing obese rat animal model based on CRISPR (clustered regularly interspaced short palindromic repeat) gene knockout technology

An animal model, gene knockout technology, applied in the field of biomedicine, can solve the problem of not being able to fully simulate the real situation of human diseases, and achieve the effect of deepening understanding

Inactive Publication Date: 2014-03-05
SUZHOU TONGSHAN BIO TECH
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Although the study of transgenic mice plays an important role in the study of

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Embodiment 1

[0031] A method for establishing an animal model of obesity rats based on CRISPR gene knockout technology, comprising the following steps:

[0032] (1) Establish a Lep / Lepr gene knockout rat model;

[0033] (2) Identification and correlation analysis of rat animal models of obesity;

[0034] (3) Evaluation of energy metabolism and body fat percentage in rat animal models of obesity.

[0035] The establishment of the Lep / Lepr gene knockout rat model specifically includes the following steps:

[0036] 1) Construction of CRISPR targeted modification gene carrier;

[0037] 2) Rat ovulation induction and in vitro fertilization;

[0038] 3) Microinjection of rat fertilized eggs;

[0039] 4) In vitro culture of fertilized eggs, implantation into recipients and breeding of targeted gene-modified animals.

[0040] The identification and related analysis of the obesity disease rat animal model specifically includes the following steps:

[0041] 1) DNA sequencing of the CRISPR targ...

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Abstract

The invention provides a method for establishing an obese rat animal model based on a CRISPR (clustered regularly interspaced short palindromic repeat) gene knockout technology. The method comprises the following steps: (1) establishing an Lep/Lepr gene knockout rat model; (2) carrying out the authentication and related analysis on the obese rat animal model; (3) evaluating the energy metabolism and the body fat rate of the obese rat animal model. According to the method, a CRISPR/Cas system is used for respectively or simultaneously knocking out Lep and LepR genes so as to obtain the rat model modified by a corresponding target gene, so that the understanding of gene regulation in the obesity morbidity process can be deepened, and the high-level animal model can be provided for the translational medicine and the new medicine research and development.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for establishing an animal model of obesity rats based on CRISPR gene knockout technology. Background technique [0002] With the improvement of living standards, obesity (obesity), as a chronic disease, has a sharp increase in incidence and has become a worldwide medical problem. Obesity is also closely related to diabetes, and 70% to 80% of diabetic patients over the age of 40 were obese before the disease. At the same time, obesity is also accompanied by symptoms such as hyperlipidemia, hypertension, and abnormal glucose tolerance, and has become the main cause of atherosclerosis. Leptin (Lep) is a hormone secreted by fat cells discovered in 1994. Through the Leptin receptor (Lepr) in the central nervous system, Lep can regulate various physiological functions and individual behaviors including blood sugar concentration, neuroendocrine, body weight, appetite, etc. Curren...

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Application Information

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IPC IPC(8): C12N15/85A01K67/027
Inventor 林卿范国平
Owner SUZHOU TONGSHAN BIO TECH
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