Abstract: Antisense oligonucleotides (ASOs) are a novel class of small molecule gene-targeted drugs that can bind to target mRNA. Through complementary base pairing with the target sequence， antisense oligonucleotides achieve targeted regulation of genes. With the continuous development of gene sequencing technology and molecular synthesis techniques， research and applications of ASOs in the musculoskeletal system are further advancing. This article reviews the mechanisms of ASOs in gene silencing and expression regulation， as well as their prospects in gene therapy. In addition， we evaluate the research progress and applications of ASOs in musculoskeletal diseases， while analyzing the urgent issues currently faced by this class of drugs. This comprehensive study aims to deepen our understanding of ASOs and provide valuable references for their widespread application in biomedical research and clinical settings.