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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...

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Autores principales: Soukupová, Marie, Zucchini, Silvia, Trempat, Pascal, Ingusci, Selene, Perrier-Biollay, Coline, Barbieri, Mario, Cattaneo, Stefano, Bettegazzi, Barbara, Falzoni, Simonetta, Berthommé, Hervé, Simonato, Michele
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2021
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.
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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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