Production of high-titer transmission-defective RNA virus-based episomal vector using tangential flow filtration
Production of high-titer transmission-defective RNA virus-based episomal vector using tangential flow filtration
复制标题
使用切向流过滤生产高滴度传输缺陷型 RNA 病毒附加型载体
DOI：
10.1111/1348-0421.12831
复制
发表时间：
2020
影响因子：
2.6
通讯作者：
Tomonaga K.
中科院分区：
文献类型：
--
作者：
Komatsu Y;Kakuya Y;Tomonaga K.
近年来，基于病毒载体的体内基因递送策略在遗传性疾病的治疗中取得了显著的成功。缺乏病毒糖蛋白基因的基于RNA病毒的游离型载体（Δ G-REVec）是一种非传递性基因递送系统，其能够在体外在多种细胞类型中进行长期基因表达，但由于难以产生高滴度载体，体内基因递送尚未成功。本研究表明，切向流过滤（TFF）可有效提高Δ G-REVec滴度。使用TFF浓缩和渗滤Δ G-REVec显著增加其滴度，而不损失感染活性。重要的是，颅内施用高滴度载体能够在啮齿动物脑中持续表达转基因。
In recent years, viral vector based in vivo gene delivery strategies have achieved a significant success in the treatment of genetic diseases. RNA virus‐based episomal vector lacking viral glycoprotein gene (ΔG‐REVec) is a nontransmissive gene delivery system that enables long‐term gene expression in a variety of cell types in vitro, yet in vivo gene delivery has not been successful due to the difficulty in producing high titer vector. The present study showed that tangential flow filtration (TFF) can be effectively employed to increase the titer of ΔG‐REVec. Concentration and diafiltration of ΔG‐REVec using TFF significantly increased its titer without loss of infectious activity. Importantly, intracranial administration of high titer vector enabled persistent transgene expression in rodent brain.