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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Vydané v:	Microbiology and immunology Ročník 64; číslo 9; s. 602 - 609
Hlavní autori:	Komatsu, Yumiko, Kakuya, Yoji, Tomonaga, Keizo
Médium:	Journal Article
Jazyk:	English
Vydavateľské údaje:	Australia Wiley Subscription Services, Inc 01.09.2020
Predmet:	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
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Abstract	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.
AbstractList	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. 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.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.
Author	Kakuya, Yoji Tomonaga, Keizo Komatsu, Yumiko
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Cites_doi	10.1111/1348-0421.12505 10.1073/pnas.90.17.8033 10.1038/sj.gt.3300938 10.1155/2013/312709 10.1038/mtm.2016.17 10.3389/fmicb.2019.02485 10.1002/jgm.1092 10.1002/jgm.778 10.1128/JVI.05554-11 10.1111/1348-0421.12602 10.1089/hgtb.2012.059 10.1016/j.omtm.2019.05.010 10.1016/j.jviromet.2007.01.002 10.1089/hum.2013.184 10.1038/sj.gt.3301188 10.1111/j.1439-0450.1997.tb00962.x 10.1016/j.jviromet.2004.08.017 10.1007/978-1-4939-3271-9_17 10.1038/mt.2015.187 10.1089/104303404772680029 10.1089/10430340260355347 10.1038/mt.2009.128 10.1038/gt.2015.108 10.1016/j.jviromet.2007.03.014 10.1089/10430340260355338 10.1016/j.chom.2012.04.009 10.1038/sj.gt.3302094
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SubjectTerms	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
Title	Production of high‐titer transmission‐defective RNA virus‐based episomal vector using tangential flow filtration
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