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Construction method of transgenic mouse skeletal muscular dystrophy model

A technology for transgenic mice and malnutrition, which is applied in the field of transgenic technology and the construction of animal disease models, and can solve problems such as being unable to be used as model animals.

Active Publication Date: 2019-11-26
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The muscle fiber basement membrane in collagenVIα1 knockout mice is intact, which is quite different from human Ullrich type congenital muscular dystrophy, so it cannot be used as a model animal for this type of disease

Method used

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  • Construction method of transgenic mouse skeletal muscular dystrophy model
  • Construction method of transgenic mouse skeletal muscular dystrophy model
  • Construction method of transgenic mouse skeletal muscular dystrophy model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The construction of embodiment 1 expression vector

[0019] The expression vectors pTet-On-α-actin and pTRE2-29 were constructed respectively. The expression vector pTet-On-α-actin uses pTet-On as the backbone vector, and replaces the Pcmv promoter in pTet-On with the α-actin promoter. The expression vector pTRE2-29 uses pTRE2 as the backbone vector, and the coding sequences of microRNA29a and microRNA29b1 are connected in series downstream of Pminicmv.

[0020] The nucleotide sequences of the constructed expression vectors pTet-On-α-actin and pTRE2-29 are shown in Seq ID No.1 and 2, respectively.

Embodiment 2

[0021] Example 2 Preparation of transgenic mice by pronuclear microinjection

[0022] The expression vectors constructed in Example 1 were used to prepare transgenic mice by pronuclear microinjection of fertilized eggs (Genetic transformation of mouse embryos by microinjection of purified DNA, Gordon, ProcNatl Acad Sci USA), and then by pTet-On transgenic mice and The pTRE2 transgenic mice were crossed to obtain mice that specifically express the rtTA gene in the offspring and express microRNA29a and microRNA29b1 in the presence of doxycycline, which is the transgenic mouse model of skeletal muscular dystrophy.

Embodiment 3

[0023] The verification of embodiment 3 experimental animal model

[0024] The mice in Example 2 were fed with doxycycline after birth, and the overexpression of microRNA29a and microRNA29b1 in offspring was realized after birth through the milk of the mother (see Post-transcriptional Regulation ofKeratinocyte Progenitor Cell Expansion, Differentiation and Hair FollicleRegression by miR-22, PLoS Genet, 2015), and observed and recorded the body weight, exercise capacity and respiratory status of the mice. Depend on figure 1 The results showed that the body weight of the transgenic mice was significantly lower than that of the control group (the transgenic mice containing pTet-On or pTRE2 were used as the control).

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Abstract

The present invention provides a transgenic mouse skeletal muscular dystrophy model construction method, wherein transgenic mice capable of specifically expressing microRNA29a and microRNA29b1 in muscle in an induction manner are constructed by using a Tet-on system and a transgenic technology so as to be used as the human Ullrich type congenital myodystrophy experiment animal model, wherein the represents of the constructed model are the same as the Ullrich type congenital myodystrophy. According to the present invention, the constructed experiment animal model has the serious respiratory problem during the growth process, wherein about 17% of the mice can die due to respiration disorders before ablactation, and all the transgenic mice can appear the skeletal muscle dysplasia, such that the powerful tool is provided for the treatment and the research of the human Ullrich type congenital myodystrophy.

Description

technical field [0001] The invention relates to a transgenic technology and a construction method of an animal disease model, in particular to a construction method of a transgenic mouse skeletal muscular dystrophy model. Background technique [0002] Animal models of human diseases refer to animals with simulated performance of human diseases established in various medical scientific researches. In the study of human diseases, suitable disease animal models provide an important theoretical basis for the study of pathogenic mechanisms. The use of experimental animal models has promoted the accumulation of clinical experience and drug development, while avoiding the risks and ethical issues caused by human experiments. Experimental animal models are divided into spontaneous animal models and induced animal models. Among them, the experimental conditions of the induced animal model are easy to control, the repeatability is good, and a large number of disease models can be in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85A01K67/027
Inventor 孟庆勇刘春城王萌张阔李宁
Owner CHINA AGRI UNIV