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Construction method and application of obese type II diabetes mellitus zebrafish model

A construction method and zebrafish technology, applied in the field of animal models, can solve the problems of little research on obese type II diabetes zebrafish models, and achieve the effect of fast and efficient construction methods, stable model characteristics and genetic traits

Active Publication Date: 2021-10-12
WENZHOU UNIVERSITY
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Problems solved by technology

[0006] However, in the current research, there are few studies on the establishment of obese type II diabetes zebrafish models by gene knockout.

Method used

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  • Construction method and application of obese type II diabetes mellitus zebrafish model
  • Construction method and application of obese type II diabetes mellitus zebrafish model

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Experimental program
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Effect test

Embodiment 1

[0040] Embodiment 1: a kind of construction method of obese type II diabetes zebrafish model, its construction steps are as follows:

[0041] 1) Obtain wild-type zebrafish eggs:

[0042] After the light is turned on, the wild-type zebrafish that are normally cultured are placed in the zebrafish mating tank according to the ratio of 1:1 or 1:2, and the male and female are placed on both sides of the baffle, and the baffle is removed after standing for 2 hours. After one hour, collect the fertilized eggs that sink to the bottom of the cylinder in time;

[0043] 2) Construction of gene knockout zebrafish:

[0044] ①The pCS2-Cas9 plasmid was digested with XbaⅠ for linearization, and the digested product was purified and transcribed into Cas9 mRNA in vitro and stored at -80°C;

[0045] ② Use the CHOPCHOP online website (https: / / chopchop.rc.fas.harvard.edu / ) to select candidate Cas9 target sites of the mc4r gene, and synthesize upstream primers and downstream primers respectively,...

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Abstract

The invention discloses a construction method and application of an obese type II diabetes mellitus zebrafish model. The obese type II diabetes mellitus zebrafish model with stable hereditary characters is constructed by means of knocking out mc4r genes, the constructed zebrafish model has symptoms of obesity and type II diabetes mellitus at the same time and can stably inherit clinical symptoms of obesity and type II diabetes mellitus, an animal model with a stable source can be obtained by natural breeding of animals through one-time construction, and compared with a simple physiological inducible model, the construction method is rapid and efficient, and the experimental efficiency of the animal model can be effectively improved and the experimental cost can be effectively reduced.

Description

technical field [0001] The invention relates to the technical field of animal models, in particular to a construction method and application of an obese type II diabetes zebrafish model. Background technique [0002] With the improvement of daily life, obesity has become a serious problem endangering human health. Obesity is a metabolic syndrome caused by excessive growth of adipose tissue caused by excessive energy intake. It can not only lead to insulin resistance, but also lead to type 2 diabetes in patients. Therefore, many obese patients are also type 2 diabetes patients. [0003] In the past few years, the animal models used to develop drug evaluation models for the clinical treatment of obesity and type II diabetes and to study their pathogenesis are usually rodents such as rabbits and mice, and the common method used to construct such animal models is to use wild-type Animals are induced by high-sugar and high-fat feeding, but there are problems such as long model c...

Claims

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Application Information

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IPC IPC(8): A01K67/027C12N15/85C12N15/12
CPCA01K67/0276C12N15/8509C07K14/461A01K2227/40A01K2267/0362A01K2217/075
Inventor 金立波彭仁义李瑶琦孙达林苏娥嵇豪
Owner WENZHOU UNIVERSITY
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