Degradation and Stable Maintenance of AAV Inverted Terminal Repeats in E. coli

Abstract

Current plasmid propagation compromises large inverted repeats, like inverted terminal repeats (ITRs) of adeno-associated virus (AAV). Direct long-read sequencing analyses upon varying culture conditions and strains revealed ITR instability mechanisms, which diminished in absence of SbcC or at elevated growth temperatures (e.g. 42 {degrees}C), with a combination being optimal. Resulting full ITRs improved rAAV yield and purity. The findings advance plasmid biology, cloneable sequences, and therapeutic AAV manufacturing.