HEK293 Transcriptome during rAAV production

The use of gene therapies based on recombinant adeno-associated viruses (rAAVs) has increased rapidly, so there is a growing need for more efficient rAAV manufacturing methods. Making viruses puts a lot of stress on the cells used to produce them, requiring a lot of energy and cellular resources. Because of this, virus production depends heavily on the condition and behavior of the host cells. To better understand this, researchers used transcriptomics, a method that studies gene activity, to find important changes in cell behavior that support rAAV production. The study looked at two different cell lines grown in their usual environments, comparing cells making the virus with those that were not, over time, using human embryonic kidney (HEK293) cells. The results showed that certain immune system pathways in the host cells were highly active during virus production. These included RIG-I-like receptor signaling, Toll-like receptor signaling, cytosolic DNA sensing, and JAK-STAT signaling. The cells also showed signs of stress, like stress in the endoplasmic reticulum, increased autophagy, and apoptosis. On the other hand, processes like fatty acid metabolism and the transport of neutral amino acids were reduced in the later stages of virus production. This analysis shows common patterns across different cell lines and provides useful information for future research aimed at making rAAV production more efficient.