Liver fibrosis is a critical pathological stage in the progression of various chronic liver diseases to liver cirrhosis and liver cancer, and it is characterized by the activation of hepatic stellate cells and excessive deposition of extracellular matrix. There are currently limited clinical treatment methods for liver fibrosis, and gene therapy has attracted increasing attention as a promising intervention strategy. Owing to its advantages of low immunogenicity, strong tissue tropism, and long-term gene expression, adeno-associated virus (AAV) has become an ideal vector for gene therapy. This article systematically reviews the structural and functional features of AAV, the tissue-targeting properties of different serotypes, and the methods for preparation and purification of recombinant AAV and focuses on the advances in the application of AAV-mediated gene overexpression, gene silencing, gene editing, and cellular reprogramming in mechanism research and intervention of liver fibrosis, as well as the challenges in clinical translation and corresponding strategies. AAV-mediated gene therapy is expected to provide novel tools and strategies for the precise treatment and mechanism study of liver fibrosis.