Abstract: Antisense oligonucleotides(ASOs) are a novel class of small molecule gene-targeted drugs that can bind with 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 have further advanced. This article reviews the mechanisms of ASOs in gene silencing and expression regulation, as well as their prospects in gene therapy. It also evaluates the research progress and applications of ASOs in musculoskeletal diseases and analyzes the urgent issues currently faced by this class of drugs. This comprehensive study aims to deepen our understanding of ASOs and provide valuable reference for their widespread application in biomedical research and clinical settings.