The Construction method and application of type ii animal model of mucopolysaccharide storage disease

Abstract

This invention provides a method for constructing type ii animal model of mucopolysaccharide storage disease, which comprises synthesizing the oligonucleotide strand and annealing annealing the pUC57 T7 SgRNA with it, then connect to gain a sgRNA expression vector according to the mouse iDS gene design target; to obtain a mouse fertilized egg; mix the transferred Cas9mRNA and sgRNA to obtain an RNA mixture, microinject the RNA mixture to the cytoplasm of that fertilized egg, then implant the it into the female mice to get a F0 generation mice; extract the genomic DNA of them and use them as a template to carry out PCR reaction. The product was digested and the mutant ones were selected to determine Founder Mice; Founder Mice and those wild male mice were mated to obtain F1 mice. The invention also provides an animal model constructed by this method and the usage of the model in the study of mucopolysaccharide sequestration type ii.

Description

technical field [0001] The invention belongs to the technical field of medical biology, in particular, the invention relates to a method for constructing an animal model. Background technique [0002] Mouse disease models play a key role in the study of human disease pathogenesis and drug screening, especially in the evaluation of drug treatment effects and the selection of treatment methods. [0003] Gene knockout is a new type of molecular biology technology developed since the late 1980s. It is a technology to inactivate or delete specific genes in the body through a certain way. Gene knockout in the general sense is mainly to apply the principle of DNA homologous recombination to replace the target gene fragment with a designed homologous fragment, so as to achieve the purpose of gene knockout. This is a complex molecular biology technology, also known as "gene targeting" technology. So far, thousands of gene mutant mouse models have been constructed using this technolo...

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Problems solved by technology

[0005] Aiming at the complicated process, harsh conditions and huge cost in the construction of the current animal model, and the lack of researchable animal models in the current mucopolysaccharidosis type II research, the present invention provides a mucopolysaccharidosis type II Method for constructing animal model, animal model obtained by the method, and use of the animal model in the study of mucopolysaccharidosis type II

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