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Improvement of HSV-1 based amplicon vectors for a safe and long-lasting gene therapy in non-replicating cells
A key factor for developing gene therapy strategies for neurological disorders is the availability of suitable vectors. Currently, the most advanced are adeno-associated vectors that, while being safe and ensuring long-lasting transgene expression, have a very limited cargo capacity. In contrast, he...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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American Society of Gene & Cell Therapy
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8044385/ https://www.ncbi.nlm.nih.gov/pubmed/33869657 http://dx.doi.org/10.1016/j.omtm.2021.03.020 |
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author | Soukupová, Marie Zucchini, Silvia Trempat, Pascal Ingusci, Selene Perrier-Biollay, Coline Barbieri, Mario Cattaneo, Stefano Bettegazzi, Barbara Falzoni, Simonetta Berthommé, Hervé Simonato, Michele |
author_facet | Soukupová, Marie Zucchini, Silvia Trempat, Pascal Ingusci, Selene Perrier-Biollay, Coline Barbieri, Mario Cattaneo, Stefano Bettegazzi, Barbara Falzoni, Simonetta Berthommé, Hervé Simonato, Michele |
author_sort | Soukupová, Marie |
collection | PubMed |
description | A key factor for developing gene therapy strategies for neurological disorders is the availability of suitable vectors. Currently, the most advanced are adeno-associated vectors that, while being safe and ensuring long-lasting transgene expression, have a very limited cargo capacity. In contrast, herpes simplex virus-based amplicon vectors can host huge amounts of foreign DNA, but concerns exist about their safety and ability to express transgenes long-term. We aimed at modulating and prolonging amplicon-induced transgene expression kinetics in vivo using different promoters and preventing transgene silencing. To pursue the latter, we deleted bacterial DNA sequences derived from vector construction and shielded the transgene cassette using AT-rich and insulator-like sequences (SAm technology). We employed luciferase and GFP as reporter genes. To determine transgene expression kinetics, we injected vectors in the hippocampus of mice that were longitudinally scanned for bioluminescence for 6 months. To evaluate safety, we analyzed multiple markers of damage and performed patch clamp electrophysiology experiments. All vectors proved safe, and we managed to modulate the duration of transgene expression, up to obtaining a stable, long-lasting expression using the SAm technology. Therefore, these amplicon vectors represent a flexible, efficient, and safe tool for gene delivery in the brain. |
format | Online Article Text |
id | pubmed-8044385 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-80443852021-04-16 Improvement of HSV-1 based amplicon vectors for a safe and long-lasting gene therapy in non-replicating cells Soukupová, Marie Zucchini, Silvia Trempat, Pascal Ingusci, Selene Perrier-Biollay, Coline Barbieri, Mario Cattaneo, Stefano Bettegazzi, Barbara Falzoni, Simonetta Berthommé, Hervé Simonato, Michele Mol Ther Methods Clin Dev Original Article A key factor for developing gene therapy strategies for neurological disorders is the availability of suitable vectors. Currently, the most advanced are adeno-associated vectors that, while being safe and ensuring long-lasting transgene expression, have a very limited cargo capacity. In contrast, herpes simplex virus-based amplicon vectors can host huge amounts of foreign DNA, but concerns exist about their safety and ability to express transgenes long-term. We aimed at modulating and prolonging amplicon-induced transgene expression kinetics in vivo using different promoters and preventing transgene silencing. To pursue the latter, we deleted bacterial DNA sequences derived from vector construction and shielded the transgene cassette using AT-rich and insulator-like sequences (SAm technology). We employed luciferase and GFP as reporter genes. To determine transgene expression kinetics, we injected vectors in the hippocampus of mice that were longitudinally scanned for bioluminescence for 6 months. To evaluate safety, we analyzed multiple markers of damage and performed patch clamp electrophysiology experiments. All vectors proved safe, and we managed to modulate the duration of transgene expression, up to obtaining a stable, long-lasting expression using the SAm technology. Therefore, these amplicon vectors represent a flexible, efficient, and safe tool for gene delivery in the brain. American Society of Gene & Cell Therapy 2021-03-29 /pmc/articles/PMC8044385/ /pubmed/33869657 http://dx.doi.org/10.1016/j.omtm.2021.03.020 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Soukupová, Marie Zucchini, Silvia Trempat, Pascal Ingusci, Selene Perrier-Biollay, Coline Barbieri, Mario Cattaneo, Stefano Bettegazzi, Barbara Falzoni, Simonetta Berthommé, Hervé Simonato, Michele Improvement of HSV-1 based amplicon vectors for a safe and long-lasting gene therapy in non-replicating cells |
title | Improvement of HSV-1 based amplicon vectors for a safe and long-lasting gene therapy in non-replicating cells |
title_full | Improvement of HSV-1 based amplicon vectors for a safe and long-lasting gene therapy in non-replicating cells |
title_fullStr | Improvement of HSV-1 based amplicon vectors for a safe and long-lasting gene therapy in non-replicating cells |
title_full_unstemmed | Improvement of HSV-1 based amplicon vectors for a safe and long-lasting gene therapy in non-replicating cells |
title_short | Improvement of HSV-1 based amplicon vectors for a safe and long-lasting gene therapy in non-replicating cells |
title_sort | improvement of hsv-1 based amplicon vectors for a safe and long-lasting gene therapy in non-replicating cells |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8044385/ https://www.ncbi.nlm.nih.gov/pubmed/33869657 http://dx.doi.org/10.1016/j.omtm.2021.03.020 |
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