Adeno-associated virus vector system controlling capsid expression improves viral quantity and quality

Kenji Ohba; Yoshihide Sehara; Tatsuji Enoki; Junichi Mineno; Keiya Ozawa; Hiroaki Mizukami

doi:10.1016/j.isci.2023.106487

Adeno-associated virus vector system controlling capsid expression improves viral quantity and quality

iScience. 2023 Mar 23;26(4):106487. doi: 10.1016/j.isci.2023.106487. eCollection 2023 Apr 21.

Authors

Kenji Ohba¹, Yoshihide Sehara¹, Tatsuji Enoki², Junichi Mineno², Keiya Ozawa^{1

3}, Hiroaki Mizukami¹

Affiliations

¹ Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi 329-0498, Japan.
² CDM Center, TAKARA Bio Inc., Kusatsu, Shiga 525-0058, Japan.
³ Department of Immuno-Gene & Cell Therapy (Takara Bio), Jichi Medical University, Shimotsuke, Tochigi 329-0498, Japan.

Abstract

Adeno-associated virus (AAV) vectors are promising tools for gene therapy. The current AAV vector system produces an abundance of empty capsids that are eliminated before clinical use, leading to increased costs for gene therapy. In the present study, we established an AAV production system that regulates the timing of capsid expression using a tetracycline-dependent promoter. Tetracycline-regulating capsid expression increased viral yield and reduced empty capsids in various serotypes without altering AAV vector infectivity in vitro and in vivo. The replicase expression pattern change observed in the developed AAV vector system improved viral quantity and quality, whereas timing control of capsid expression reduced empty capsids. These findings provide a new perspective on the development of AAV vector production systems in gene therapy.

Keywords: Biotechnology; Virology.