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Assembly and Functional Analysis of an S/MAR Based Episome with the Cystic Fibrosis Transmembrane Conductance Regulator Gene
Improving the efficacy of gene therapy vectors is still an important goal toward the development of safe and efficient gene therapy treatments. S/MAR (scaffold/matrix attached region)-based vectors are maintained extra-chromosomally in numerous cell types, which is similar to viral-based vectors. Ad...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5979583/ https://www.ncbi.nlm.nih.gov/pubmed/29673202 http://dx.doi.org/10.3390/ijms19041220 |
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author | De Rocco, Davide Pompili, Barbara Castellani, Stefano Morini, Elena Cavinato, Luca Cimino, Giuseppe Mariggiò, Maria A Guarnieri, Simone Conese, Massimo Del Porto, Paola Ascenzioni, Fiorentina |
author_facet | De Rocco, Davide Pompili, Barbara Castellani, Stefano Morini, Elena Cavinato, Luca Cimino, Giuseppe Mariggiò, Maria A Guarnieri, Simone Conese, Massimo Del Porto, Paola Ascenzioni, Fiorentina |
author_sort | De Rocco, Davide |
collection | PubMed |
description | Improving the efficacy of gene therapy vectors is still an important goal toward the development of safe and efficient gene therapy treatments. S/MAR (scaffold/matrix attached region)-based vectors are maintained extra-chromosomally in numerous cell types, which is similar to viral-based vectors. Additionally, when established as an episome, they show a very high mitotic stability. In the present study we tested the idea that addition of an S/MAR element to a CFTR (cystic fibrosis transmembrane conductance regulator) expression vector, may allow the establishment of a CFTR episome in bronchial epithelial cells. Starting from the observation that the S/MAR vector pEPI-EGFP (enhanced green fluorescence protein) is maintained as an episome in human bronchial epithelial cells, we assembled the CFTR vector pBQ-S/MAR. This vector, transfected in bronchial epithelial cells with mutated CFTR, supported long term wt CFTR expression and activity, which in turn positively impacted on the assembly of tight junctions in polarized epithelial cells. Additionally, the recovery of intact pBQ-S/MAR, but not the parental vector lacking the S/MAR element, from transfected cells after extensive proliferation, strongly suggested that pBQ-S/MAR was established as an episome. These results add a new element, the S/MAR, that can be considered to improve the persistence and safety of gene therapy vectors for cystic fibrosis pulmonary disease. |
format | Online Article Text |
id | pubmed-5979583 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-59795832018-06-10 Assembly and Functional Analysis of an S/MAR Based Episome with the Cystic Fibrosis Transmembrane Conductance Regulator Gene De Rocco, Davide Pompili, Barbara Castellani, Stefano Morini, Elena Cavinato, Luca Cimino, Giuseppe Mariggiò, Maria A Guarnieri, Simone Conese, Massimo Del Porto, Paola Ascenzioni, Fiorentina Int J Mol Sci Article Improving the efficacy of gene therapy vectors is still an important goal toward the development of safe and efficient gene therapy treatments. S/MAR (scaffold/matrix attached region)-based vectors are maintained extra-chromosomally in numerous cell types, which is similar to viral-based vectors. Additionally, when established as an episome, they show a very high mitotic stability. In the present study we tested the idea that addition of an S/MAR element to a CFTR (cystic fibrosis transmembrane conductance regulator) expression vector, may allow the establishment of a CFTR episome in bronchial epithelial cells. Starting from the observation that the S/MAR vector pEPI-EGFP (enhanced green fluorescence protein) is maintained as an episome in human bronchial epithelial cells, we assembled the CFTR vector pBQ-S/MAR. This vector, transfected in bronchial epithelial cells with mutated CFTR, supported long term wt CFTR expression and activity, which in turn positively impacted on the assembly of tight junctions in polarized epithelial cells. Additionally, the recovery of intact pBQ-S/MAR, but not the parental vector lacking the S/MAR element, from transfected cells after extensive proliferation, strongly suggested that pBQ-S/MAR was established as an episome. These results add a new element, the S/MAR, that can be considered to improve the persistence and safety of gene therapy vectors for cystic fibrosis pulmonary disease. MDPI 2018-04-17 /pmc/articles/PMC5979583/ /pubmed/29673202 http://dx.doi.org/10.3390/ijms19041220 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article De Rocco, Davide Pompili, Barbara Castellani, Stefano Morini, Elena Cavinato, Luca Cimino, Giuseppe Mariggiò, Maria A Guarnieri, Simone Conese, Massimo Del Porto, Paola Ascenzioni, Fiorentina Assembly and Functional Analysis of an S/MAR Based Episome with the Cystic Fibrosis Transmembrane Conductance Regulator Gene |
title | Assembly and Functional Analysis of an S/MAR Based Episome with the Cystic Fibrosis Transmembrane Conductance Regulator Gene |
title_full | Assembly and Functional Analysis of an S/MAR Based Episome with the Cystic Fibrosis Transmembrane Conductance Regulator Gene |
title_fullStr | Assembly and Functional Analysis of an S/MAR Based Episome with the Cystic Fibrosis Transmembrane Conductance Regulator Gene |
title_full_unstemmed | Assembly and Functional Analysis of an S/MAR Based Episome with the Cystic Fibrosis Transmembrane Conductance Regulator Gene |
title_short | Assembly and Functional Analysis of an S/MAR Based Episome with the Cystic Fibrosis Transmembrane Conductance Regulator Gene |
title_sort | assembly and functional analysis of an s/mar based episome with the cystic fibrosis transmembrane conductance regulator gene |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5979583/ https://www.ncbi.nlm.nih.gov/pubmed/29673202 http://dx.doi.org/10.3390/ijms19041220 |
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