Abstract

The purpose of this study was to simulate muscular dystrophy disease by constructing a laminin-alpha 2 zebrafish model, and to study the changes of muscle transcriptome using transcriptomic methods. Using clustered regularly interspaced short palindromic repeats associated protein 9 gene editing technology; the zebrafish laminin-alpha 2 gene was knocked out or mutated. First, the appropriate guide RNA sequence was designed, and then the guide RNA and clustered regularly interspaced short palindromic repeats associated protein 9 proteins were synthesized and injected into zebrafish embryos. The lamininalpha 2 model fish is screened and identified by detecting the mutation of DNA or messenger RNA in the progeny fish genome. The muscle phenotype identification results of zebrafish model 4 showed that the muscle phenotypes of normal and unable swimming individuals in laminin-alpha 2 knockout group were abnormal, while the muscle phenotypes of clustered regularly interspaced short palindromic repeats associated protein 9 control group were not. The muscular dystrophy characteristics of laminin-alpha 2 model fish were confirmed. Functional enrichment and pathway analysis of differentially expressed genes were performed to further reveal the biological processes and regulatory pathways associated with muscular dystrophy in laminin-alpha 2 model fish. Transcriptome sequencing analysis revealed the difference in transcriptome expression between the muscle tissue of laminin-alpha 2 zebrafish model and wild-type zebrafish. These differentially expressed genes are involved in biological processes such as muscle development, metabolism and signal transduction, providing important clues for further understanding of the pathogenesis of muscular dystrophy. Through functional enrichment and pathway analysis, the key signaling pathways and molecular mechanisms involved in the pathogenesis of muscular dystrophy were identified. These findings provide theoretical basis for the discovery of new therapeutic targets and formulation of corresponding therapeutic strategies, and provide new ideas and methods for the treatment of muscle diseases.