The blood-brain barrier (BBB) is a formidable barrier that restricts the entry of substances into the brain, complicating the study of brain function and the treatment of neurological conditions. Traditional methods of delivering genes from the periphery to the central nervous system (CNS) using adeno-associated viruses (AAVs) often require high doses, which can trigger immune responses and hepatotoxicity. Here, we developed a new AAV variant named AAVhu.32-PLUS based on a rational strategy. Following intravenous injection, AAVhu.32-PLUS can cross the BBB and exhibits higher efficiency and specificity in transducing neurons and significantly reduced hepatotropism compared to the extensively used AAV-PHP.eB. Furthermore, through in vitro cell experiments, we identified that AAVhu.32-PLUS may rely on the LY6A receptor for crossing the BBB. Finally, our research indicates that AAVhu.32-PLUS, while having lower transduction efficiency in astrocytes compared to AAV-PHP.eB, is still capable of efficiently transducing glioblastoma after intravenous injection. These properties make AAVhu.32-PLUS a promising tool for neuroscience research and targeted therapies of brain disease.