Adeno-associated viruses (AAVs) are small single-stranded DNA viruses, with a genome encoding three proteins, Cap, Rep, and Aap. AAVs are currently the most studied vector for gene therapy due to their defined tissue tropism, small size, and inability to cause disease. The AAV capsid, comprised of three Cap isoforms, is an essential component of AAV function as a gene therapy vector. The AAV capsid binds to receptors for internalization via endocytosis, then undergoes conformational changes for endosome escape, prior to nuclear entry and subsequent genome release inside the nucleus. Despite these important roles for gene therapy, the mechanisms underlying nuclear translocation remain largely unresolved. A novel sequence form of porcine AAV Po1 was discovered in 2009 which raised the possibility of designing a hybrid gene therapy vector; however little is known about this new variant and in particular the role of the capsid in viral translocation to the nucleus. Initial computational analyses revealed a nuclear localization sequence (NLS) motif within the porcine AAV capsid. This NLS region was expressed, purified and complexed with importin-alpha (IMPα) and a structure was solved at 2.2Å resolution. Key binding determinants responsible for porcine AAV capsid interaction with IMPα were identified. Overall, this is the first time that a suspected NLS sequence of an AAV capsid protein in complex with host nuclear transport receptor has been determined structurally. Our results support the findings that AAV capsids enter the nucleus through binding the nuclear import adapter, IMPα.