Assembly and Functional Analysis of an S/MAR Based Episome with the Cystic Fibrosis Transmembrane Conductance Regulator Gene
| Autor: | Fiorentina Ascenzioni, Stefano Castellani, Maria A. Mariggiò, Simone Guarnieri, Paola Del Porto, Davide De Rocco, Barbara Pompili, Giuseppe Cimino, Elena Morini, Massimo Conese, Luca Cavinato |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Genetic enhancement Genetic Vectors Cystic Fibrosis Transmembrane Conductance Regulator Bronchi Respiratory Mucosa Vectors in gene therapy Transfection Cystic fibrosis Article Catalysis Cell Line S/MAR cystic fibrosis lcsh:Chemistry Inorganic Chemistry 03 medical and health sciences 0302 clinical medicine medicine Humans Vector (molecular biology) CFTR Physical and Theoretical Chemistry lcsh:QH301-705.5 Molecular Biology Mitosis Spectroscopy Expression vector biology episome gene therapy Organic Chemistry Epithelial Cells Genetic Therapy General Medicine medicine.disease Cystic fibrosis transmembrane conductance regulator Computer Science Applications Cell biology catalysis molecular biology spectroscopy computer science applications computer vision and pattern recognition physical and theoretical chemistry organic chemistry inorganic chemistry 030104 developmental biology lcsh:Biology (General) lcsh:QD1-999 030220 oncology & carcinogenesis biology.protein Plasmids |
| Zdroj: | International Journal of Molecular Sciences; Volume 19; Issue 4; Pages: 1220 International Journal of Molecular Sciences, Vol 19, Iss 4, p 1220 (2018) International Journal of Molecular Sciences |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms19041220 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|