Cellular antisilencing elements support transgene expression from herpes simplex virus vectors in the absence of immediate early gene expression

F Han, Y Miyagawa, G Verlengia, S Ingusci, M Soukupova, M Simonato, JC Glorioso, JB Cohen

Research output: Contribution to journalArticle

Abstract

Inactivation of all herpes simplex virus (HSV) immediate early (IE) genes to eliminate vector cytotoxicity results in rapid silencing of the viral genome, similar to the establishment of HSV latency. We recently reported that silencing of a nonviral reporter cassette could be overcome in nonneuronal cells by positioning the cassette in the viral latency (LAT) locus between resident chromatin boundary elements. Here, we tested the abilities of the chicken hypersensitive site 4 insulator and the human ubiquitous chromatin opening element A2UCOE to promote transgene expression from an IE-gene-inactivated HSV vector. We found that A2UCOE was particularly active in nonneuronal cells and reduced reporter promoter occupancy by a repressive histone mark. We determined whether multiple transgenes could be expressed under the control of different promoters from different loci of the same virus. The results showed abundant coexpression of LAT-embedded and A2UCOEflanked genes in nonneuronal cells. In addition, a third reporter gene without known protective elements was active in cultured rat sensory neurons. These findings indicate that cellular antisilencing sequences can contribute to the expression of multiple genes from separate promoters in fully IE gene-disabled HSV vectors, providing an opportunity for therapeutic applications requiring mutually independent expression of different gene products from a single vector. © 2018 American Society for Microbiology.
Original languageEnglish
Article numbere00536-18
JournalJournal of Virology
Volume92
Issue number17
DOIs
Publication statusPublished - 2018

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herpes simplex
Immediate-Early Genes
Simplexvirus
Transgenes
transgenes
Virus Latency
Gene Expression
gene expression
viruses
Chromatin
promoter regions
genes
Histone Code
Insulator Elements
chromatin
Viral Genome
Sensory Receptor Cells
Microbiology
Reporter Genes
loci

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Cellular antisilencing elements support transgene expression from herpes simplex virus vectors in the absence of immediate early gene expression. / Han, F; Miyagawa, Y; Verlengia, G; Ingusci, S; Soukupova, M; Simonato, M; Glorioso, JC; Cohen, JB.

In: Journal of Virology, Vol. 92, No. 17, e00536-18, 2018.

Research output: Contribution to journalArticle

Han, F, Miyagawa, Y, Verlengia, G, Ingusci, S, Soukupova, M, Simonato, M, Glorioso, JC & Cohen, JB 2018, 'Cellular antisilencing elements support transgene expression from herpes simplex virus vectors in the absence of immediate early gene expression', Journal of Virology, vol. 92, no. 17, e00536-18. https://doi.org/10.1128/JVI.00536-18
Han, F ; Miyagawa, Y ; Verlengia, G ; Ingusci, S ; Soukupova, M ; Simonato, M ; Glorioso, JC ; Cohen, JB. / Cellular antisilencing elements support transgene expression from herpes simplex virus vectors in the absence of immediate early gene expression. In: Journal of Virology. 2018 ; Vol. 92, No. 17.
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AU - Miyagawa, Y

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AU - Soukupova, M

AU - Simonato, M

AU - Glorioso, JC

AU - Cohen, JB

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AB - Inactivation of all herpes simplex virus (HSV) immediate early (IE) genes to eliminate vector cytotoxicity results in rapid silencing of the viral genome, similar to the establishment of HSV latency. We recently reported that silencing of a nonviral reporter cassette could be overcome in nonneuronal cells by positioning the cassette in the viral latency (LAT) locus between resident chromatin boundary elements. Here, we tested the abilities of the chicken hypersensitive site 4 insulator and the human ubiquitous chromatin opening element A2UCOE to promote transgene expression from an IE-gene-inactivated HSV vector. We found that A2UCOE was particularly active in nonneuronal cells and reduced reporter promoter occupancy by a repressive histone mark. We determined whether multiple transgenes could be expressed under the control of different promoters from different loci of the same virus. The results showed abundant coexpression of LAT-embedded and A2UCOEflanked genes in nonneuronal cells. In addition, a third reporter gene without known protective elements was active in cultured rat sensory neurons. These findings indicate that cellular antisilencing sequences can contribute to the expression of multiple genes from separate promoters in fully IE gene-disabled HSV vectors, providing an opportunity for therapeutic applications requiring mutually independent expression of different gene products from a single vector. © 2018 American Society for Microbiology.

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