Adeno-associated viruses (AAV) contain minimal viral proteins necessary for their replication. During virus assembly, AAV acquire, inherently and submissively, various cellular proteins. Our previous studies identified the association of AAV vectors with the DNA binding protein nucleophosmin (NPM1). Nucleophosmin has been reported to enhance AAV infection by mobilizing AAV capsids into and out of the nucleolus, indicating the importance of NPM1 in the AAV life cycle; however the role of NPM1 in AAV production remains unknown. In this study, we systematically investigated NPM1 function on AAV production using NPM1 knockdown cells and revealing for the first time the presence of G-quadruplex DNA sequences (GQRS) in the AAV genome, the synergistic NPM1-GQRS function in AAV production and the significant enhancement of NPM1 gene knockdown on AAV vector production. Understanding the role of cellular proteins in the AAV life cycle will greatly facilitate high titre production of AAV vectors for clinical use.

腺相关病毒（AAV）包含复制所需的最小病毒蛋白。在病毒组装过程中，AAV固有地和顺从地获取各种细胞蛋白。我们以前的研究确定了AAV载体与DNA结合蛋白核磷酸蛋白（NPM1）的关联。据报道，核蛋白可通过动员AAV衣壳进入和移出核仁来增强AAV感染，这表明NPM1在AAV生命周期中的重要性。然而，NPM1在AAV生产中的作用仍然未知。在这项研究中，我们系统地研究了NPM1在使用NPM1敲低细胞的AAV生产中的功能，并首次揭示了AAV基因组中G-四链体DNA序列（GQRS）的存在，NPM1-GQRS在AAV生产中的协同作用以及NPM1基因敲低对AAV载体生产的显着增强。了解细胞蛋白在AAV生命周期中的作用将大大促进AAV载体在临床上的高滴度生产。