Premium
Production of high‐titer transmission‐defective RNA virus‐based episomal vector using tangential flow filtration
Author(s) -
Komatsu Yumiko,
Kakuya Yoji,
Tomonaga Keizo
Publication year - 2020
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/1348-0421.12831
Subject(s) - biology , titer , gene delivery , vector (molecular biology) , virology , transgene , virus , viral vector , gene , in vivo , rna , gene expression , genetic enhancement , microbiology and biotechnology , recombinant dna , genetics
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.