Production of rAAV by plasmid transfection induces antiviral and inflammatory responses in suspension HEK293

To understand the response of host cells to rAAV vector production via plasmid transfection, samples were collected throughout the course of three 50 L production batches and analyzed by bulk RNA sequencing of polyadenylated transcripts (RNA-seq). Gene ontology analysis determined that upregulated pathways included inflammatory and antiviral responses. Systematic analyses of the cellular transcriptional response to rAAV production indicates that these host cells are not passively supporting vector manufacture, and therefore may illuminate genes and pathways that influence rAAV production, thereby enabling the rational design of next-generation manufacturing platforms to support safe, effective, and affordable AAV-based gene therapies. Overall design: Samples were collected immediately prior to transfection (0 h) and at 3, 6, 12, 24, 36, 48, and 72 hours post transfection (hpt).