Cargando…
In vitro Inactivation of Latent HSV by Targeted Mutagenesis Using an HSV-specific Homing Endonuclease
Following acute infection, herpes simplex virus (HSV) establishes latency in sensory neurons, from which it can reactivate and cause recurrent disease. Available antiviral therapies do not affect latent viral genomes; therefore, they do not prevent reactivation following therapy cessation. One possi...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2014
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3951911/ https://www.ncbi.nlm.nih.gov/pubmed/24496438 http://dx.doi.org/10.1038/mtna.2013.75 |
_version_ | 1782307151530688512 |
---|---|
author | Aubert, Martine Boyle, Nicole M Stone, Daniel Stensland, Laurence Huang, Meei-Li Magaret, Amalia S Galetto, Roman Rawlings, David J Scharenberg, Andrew M Jerome, Keith R |
author_facet | Aubert, Martine Boyle, Nicole M Stone, Daniel Stensland, Laurence Huang, Meei-Li Magaret, Amalia S Galetto, Roman Rawlings, David J Scharenberg, Andrew M Jerome, Keith R |
author_sort | Aubert, Martine |
collection | PubMed |
description | Following acute infection, herpes simplex virus (HSV) establishes latency in sensory neurons, from which it can reactivate and cause recurrent disease. Available antiviral therapies do not affect latent viral genomes; therefore, they do not prevent reactivation following therapy cessation. One possible curative approach involves the introduction of DNA double strand breaks in latent HSV genomes by rare-cutting endonucleases, leading to mutagenesis of essential viral genes. We tested this approach in an in vitro HSV latency model using the engineered homing endonuclease (HE) HSV1m5, which recognizes a sequence in the HSV-1 gene U(L)19, encoding the virion protein VP5. Coexpression of the 3′-exonuclease Trex2 with HEs increased HE-mediated mutagenesis frequencies up to sixfold. Following HSV1m5/Trex2 delivery with adeno-associated viral (AAV) vectors, the target site was mutated in latent HSV genomes with no detectable cell toxicity. Importantly, HSV production by latently infected cells after reactivation was decreased after HSV1m5/Trex2 exposure. Exposure to histone deacetylase inhibitors prior to HSV1m5/Trex2 treatment increased mutagenesis frequencies of latent HSV genomes another two- to fivefold, suggesting that chromatin modification may be a useful adjunct to gene-targeting approaches. These results support the continuing development of HEs and other nucleases (ZFNs, TALENs, CRISPRs) for cure of chronic viral infections. |
format | Online Article Text |
id | pubmed-3951911 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-39519112014-03-13 In vitro Inactivation of Latent HSV by Targeted Mutagenesis Using an HSV-specific Homing Endonuclease Aubert, Martine Boyle, Nicole M Stone, Daniel Stensland, Laurence Huang, Meei-Li Magaret, Amalia S Galetto, Roman Rawlings, David J Scharenberg, Andrew M Jerome, Keith R Mol Ther Nucleic Acids Original Article Following acute infection, herpes simplex virus (HSV) establishes latency in sensory neurons, from which it can reactivate and cause recurrent disease. Available antiviral therapies do not affect latent viral genomes; therefore, they do not prevent reactivation following therapy cessation. One possible curative approach involves the introduction of DNA double strand breaks in latent HSV genomes by rare-cutting endonucleases, leading to mutagenesis of essential viral genes. We tested this approach in an in vitro HSV latency model using the engineered homing endonuclease (HE) HSV1m5, which recognizes a sequence in the HSV-1 gene U(L)19, encoding the virion protein VP5. Coexpression of the 3′-exonuclease Trex2 with HEs increased HE-mediated mutagenesis frequencies up to sixfold. Following HSV1m5/Trex2 delivery with adeno-associated viral (AAV) vectors, the target site was mutated in latent HSV genomes with no detectable cell toxicity. Importantly, HSV production by latently infected cells after reactivation was decreased after HSV1m5/Trex2 exposure. Exposure to histone deacetylase inhibitors prior to HSV1m5/Trex2 treatment increased mutagenesis frequencies of latent HSV genomes another two- to fivefold, suggesting that chromatin modification may be a useful adjunct to gene-targeting approaches. These results support the continuing development of HEs and other nucleases (ZFNs, TALENs, CRISPRs) for cure of chronic viral infections. Nature Publishing Group 2014-02 2014-02-04 /pmc/articles/PMC3951911/ /pubmed/24496438 http://dx.doi.org/10.1038/mtna.2013.75 Text en Copyright © 2014 The American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by-nc-nd/3.0/ Molecular Therapy-Nucleic Acids is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivative Works 3.0 License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/ |
spellingShingle | Original Article Aubert, Martine Boyle, Nicole M Stone, Daniel Stensland, Laurence Huang, Meei-Li Magaret, Amalia S Galetto, Roman Rawlings, David J Scharenberg, Andrew M Jerome, Keith R In vitro Inactivation of Latent HSV by Targeted Mutagenesis Using an HSV-specific Homing Endonuclease |
title | In vitro Inactivation of Latent HSV by Targeted Mutagenesis Using an HSV-specific Homing Endonuclease |
title_full | In vitro Inactivation of Latent HSV by Targeted Mutagenesis Using an HSV-specific Homing Endonuclease |
title_fullStr | In vitro Inactivation of Latent HSV by Targeted Mutagenesis Using an HSV-specific Homing Endonuclease |
title_full_unstemmed | In vitro Inactivation of Latent HSV by Targeted Mutagenesis Using an HSV-specific Homing Endonuclease |
title_short | In vitro Inactivation of Latent HSV by Targeted Mutagenesis Using an HSV-specific Homing Endonuclease |
title_sort | in vitro inactivation of latent hsv by targeted mutagenesis using an hsv-specific homing endonuclease |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3951911/ https://www.ncbi.nlm.nih.gov/pubmed/24496438 http://dx.doi.org/10.1038/mtna.2013.75 |
work_keys_str_mv | AT aubertmartine invitroinactivationoflatenthsvbytargetedmutagenesisusinganhsvspecifichomingendonuclease AT boylenicolem invitroinactivationoflatenthsvbytargetedmutagenesisusinganhsvspecifichomingendonuclease AT stonedaniel invitroinactivationoflatenthsvbytargetedmutagenesisusinganhsvspecifichomingendonuclease AT stenslandlaurence invitroinactivationoflatenthsvbytargetedmutagenesisusinganhsvspecifichomingendonuclease AT huangmeeili invitroinactivationoflatenthsvbytargetedmutagenesisusinganhsvspecifichomingendonuclease AT magaretamalias invitroinactivationoflatenthsvbytargetedmutagenesisusinganhsvspecifichomingendonuclease AT galettoroman invitroinactivationoflatenthsvbytargetedmutagenesisusinganhsvspecifichomingendonuclease AT rawlingsdavidj invitroinactivationoflatenthsvbytargetedmutagenesisusinganhsvspecifichomingendonuclease AT scharenbergandrewm invitroinactivationoflatenthsvbytargetedmutagenesisusinganhsvspecifichomingendonuclease AT jeromekeithr invitroinactivationoflatenthsvbytargetedmutagenesisusinganhsvspecifichomingendonuclease |