Extended Data Fig. 2: The low-cis triple transfection method is more limited in packaging UGA-sup-tRNAs with augmented readthrough capabilities.
From: An engineered UGA suppressor tRNA gene for disease-agnostic AAV delivery
(a) Schematics showing the differences between standard triple transfection and low-cis triple transfection. The three plasmids (pHelper, pTrans, pCis) with the key elements involved in rAAV production are illustrated. (b) Bar graph showing the rAAV titers in the crude lysates of HEK293 cell cultures when packaging indicated pCis with various input amount. Data are normalized to the titer of rAAV9.mCherry produced with 100% pCis in parallel (defined as 100%) and presented as percentages. Each dot represents a biological repeat (n = 3). Data are presented as mean and standard deviation. (c) Western blot images showing the Cap (top), Rep (middle), and GAPDH (bottom, serving as the loading control) protein expression in HEK293 cells undergoing rAAV9 production with indicated pCis and amount. 3 independent experiments were conducted.
