Production of high‐titer transmission‐defective RNA virus‐based episomal vector using tangential flow filtration

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 i...

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Bibliographic Details
Published in:Microbiology and immunology Vol. 64; no. 9; pp. 602 - 609
Main Authors: Komatsu, Yumiko, Kakuya, Yoji, Tomonaga, Keizo
Format: Journal Article
Language:English
Published: Australia Wiley Subscription Services, Inc 01.09.2020
Subjects:
Animals
Brain - virology
Cell Line
Chlorocebus aethiops
Female
Filtration
Filtration - methods
Gene Expression
Gene transfer
Gene Transfer Techniques
Genetic Vectors - administration & dosage
Genetic Vectors - isolation & purification
HEK293 Cells
high‐titer
Humans
in vivo gene delivery
Plasmids - genetics
Plasmids - isolation & purification
Pregnancy
Rats
Rats, Inbred Lew
RNA viruses
RNA Viruses - genetics
RNA Viruses - isolation & purification
RNA virus‐based episomal vector
tangential flow filtration
Transgenes
Vero Cells
Viral Load
viral vector
in vivo gene delivery
viral vector
RNA virus-based episomal vector
tangential flow filtration
high-titer
ISSN:0385-5600, 1348-0421, 1348-0421
Online Access:Get full text
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Summary: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.
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ISSN:0385-5600
1348-0421
1348-0421
DOI:10.1111/1348-0421.12831