PROJECT SUMMARY (Core C – Viral Vector Core) The potential of recombinant adeno-associated virus vector (rAAV)-mediated delivery of HIV inhibitors with broadly neutralizing activities to HIV infection is clearly demonstrated in work from Phil Johnson, Reed Clark, and Ronald Desrosiers (Nature Medicine 2009), Alex Balazs and David Baltimore (Nature 2012), Matthew Gardner and Michael Farzan (Nature 2015). The therapeutic potential of AAV-expressed inhibitors is clearly shown in this proposal and recent published work from the Desrosiers Lab (Immunity 2019). The main objective of the Vector Core is to provide high quality single-stranded (ss) and self-complementary (sc) rAAV vectors to support the proposed studies. We will accomplish this goal through two specific aims. Aim 1 will design, create, produce, and quality control test scAAV vector lots at different scales with a variety of capsids, transgenes and expression cassettes to serve the specific needs of other investigators of this program project. 154 rhesus macaques will be enrolled for different studies over 4 years, and hundreds of mice will be used for pre-macaque evaluation. On average, we estimate that -30 vector lots will be produced annually to meet the needs of those studies. Aim will develop a novel and scalable rAAV production method for larger scale translational NHP studies and future clinical development of rAAV-based anti-HIV vaccine and therapeutics. Our current AAV production system should meet the vector needs in the early stage of this program project. However, large scale vector production may become a bottle neck for larger translational NHP studies and future clinical development. We will utilize our extensive experience in developing various vector packaging cell lines, suspension cell culture-based vector production, corresponding downstream processing and chromatography-based purification systems to develop a scalable suspension 293 cell-based production method to overcome this limitation. In doing so, we further our common goal of establishing AAV-mediated functional cures in macaques and humans.