Update publication information. Gene therapy has been thriving in recent years, as featured by general triumphs in several clinical trials to treat hemophilia. It highlights the significance of AAV mediated gene delivery to liver. AAV5 is a unique serotype with low neutralizing antibody prevalence. In this study, we generated an AAV5 mutant vector by inserting an oligopeptide into the variable region VIII of the AAV5, termed as AAVzk2, which displayed comparable yield with wild type serotypes. Mice intravenously injected with AAVzk2 demonstrated a stronger liver transduction than AAV5, which were grossly comparable with AAV8 and 9 treated mice. However, AAVzk2 negligibly transduced other tissues or organs such as heart, lung, spleen, kidney, brain and skeletal muscle, indicating a liver specific tropism. Further studies showed a superior human hepatocellular transduction of AAVzk2 to AAV5, 8 and 9, whereas the seroreactivity of AAVzk2 was as low as AAV5. Moreover, mass spectroscopy revealed great divergency in post-translational modification patterns between AAV5 and AAVzk2. Overall, we provide a novel AAV serotype that could not only facilitate a robust and specific liver gene delivery to a large population, but also offer valuable clues for future investigation of the nature of AAV5.