Preparation method of zebra fish model for teenager idiopathic scoliosis

Abstract

The invention discloses a preparation method of a zebra fish model for teenager idiopathic scoliosis. The preparation method comprises the following steps: S1, synthesizing RNP; s2, injecting RNP into a wild zebra fish embryo within 1 hour of birth to obtain an F0 embryo; s3, carrying out PCR detection on the mutation condition of the target site; s4, screening zebrafish genetic lines with stable gene knockout; s5, selecting tail-rolling malformed juvenile fishes, feeding the juvenile fishes until the juvenile fishes are adult, and regularly observing phenotypes; s6, extracting DNA from the adult zebra fish, and detecting to confirm that gene editing succeeds; and S7, confirming successful establishment of the model according to a DNA sequencing result and zebra fish lateral convex phenotype, belonging to the technical field of animal models, and starting from a positive result of clinical genetics screening, enabling the model to have the representativeness of the disease; a gene editing method is adopted, mutation is edited into a zebra fish genome, a Cas9 gene editing technology is used for experiment, the gene editing efficiency is ensured, and the positive rate of the constructed lateral convex model is high.

Description

technical field [0001] The invention belongs to the technical field of animal models, in particular to a method for preparing a zebrafish model of juvenile idiopathic scoliosis. Background technique [0002] AIS is a developmental deformity disease characterized by curvature of the 3D structure of the spine that occurs in adolescent women. It is also one of the most common skeletal system deformities in the world, with an incidence of about 1-5% in the population, of which about 10% of the patients have worsening symptoms and require clinical intervention. Currently, there is only surgical orthopaedic treatment for severe IS patients, but the surgical trauma is relatively high, the cost is high, and there are a series of complications, which have a negative impact on the physical and mental development of the patients, their families, and their families. and society has created a huge burden. Therefore, a detailed understanding of the occurrence and development mechanism of...

AI Technical Summary

Problems solved by technology

[0004] In zebrafish research, it has been found that knockout of several genes can lead to the scoliosis phenotype in zebrafish, but none of these pathogenic genes have been verified in AIS patients (that is, the association of these genes with the disease remains to be studied), This makes the correlation between these gene knockout zebrafish and human diseases greatly reduced and cannot be used for subsequent mechanism studies

[0005] Previous studies have used mice to establish scoliosis animal models, but the construction period of transgenic mice is long, the cost is high, and the manpower and material resources required for feeding are large, and follow-up studies not only need to explore the pathogenesis, but also need to be on the corresponding Experimental research with molecularly targeted therapies, which may involve editing or knocking out multiple genes

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