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bioRxiv · 10.1101/2024.06.03.597121

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

Radukic, M. T.·Le, D. T.·Krassuski, T.·Borchert, P.·Leach, D. R. F.·Müller, K. M.
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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.

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Radukic, M. T., Le, D. T., Krassuski, T., Borchert, P., Leach, D. R. F., Müller, K. M.. 2024-06-03. Degradation and Stable Maintenance of AAV Inverted Terminal Repeats in E. coli. https://doi.org/10.1101/2024.06.03.597121

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