دورية أكاديمية
Improved osteogenic vector for non-viral gene therapy
| العنوان: | Improved osteogenic vector for non-viral gene therapy |
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| المؤلفون: | ARA Hacobian, K Posa-Markaryan, S Sperger, M Stainer, D Hercher, GA Feichtinger, CMAP Schuh, H Redl |
| المصدر: | European Cells & Materials, Vol 31, Pp 191-204 (2016) |
| بيانات النشر: | Forum Multimedia Publishing LLC, 2016. |
| سنة النشر: | 2016 |
| المجموعة: | LCC:Diseases of the musculoskeletal system LCC:Orthopedic surgery |
| مصطلحات موضوعية: | Bone morphogenetic protein, gene therapy, osteogenic differentiation, bone regeneration, codon optimisation, intron, non-viral, secretion signal, Diseases of the musculoskeletal system, RC925-935, Orthopedic surgery, RD701-811 |
| الوصف: | Therapeutic compensation of deficient bone regeneration is a challenging task and a topic of on-going search for novel treatment strategies. One promising approach for improvement involves non-viral gene delivery using the bone morphogenetic protein-2 (BMP-2) gene to provide transient, local and sustained expression of the growth factor. However, since efficiency of non-viral gene delivery is low, this study focused on the improvement of a BMP-2 gene expression system, aiming for compensation of poor transfection efficiency. First, the native BMP-2 gene sequence was modified by codon optimisation and altered by inserting a highly truncated artificial intron (96 bp). Transfection of multiple cell lines and rat adipose-derived mesenchymal stem cells with plasmids harbouring the improved BMP-2 sequence led to a several fold increased expression rate and subsequent osteogenic differentiation. Additionally, comparing expression kinetics of elongation factor 1 alpha (EF1α) promoter with a state of the art CMV promoter revealed significantly higher BMP-2 expression when under the influence of the EF1α promoter. Results obtained by quantification of bone markers as well as osteogenic assays showed reduced sensitivity to promoter silencing effects of the EF1α promoter in rat adipose-derived mesenchymal stem cells. Finally, screening of several protein secretion signals using either luciferase or BMP-2 as reporter protein revealed no superior candidates for potential replacement of the native BMP-2 secretion signal. Taken together, by enhancing the exogenous BMP-2 expression system, low transfection efficiencies in therapeutic applications can be compensated, making safe non-viral systems even more suitable for tissue regeneration approaches. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 1473-2262 |
| Relation: | http://www.ecmjournal.org/papers/vol031/pdf/v031a13.pdf; https://doaj.org/toc/1473-2262 |
| DOI: | 10.22203/eCM.v031a13 |
| URL الوصول: | https://doaj.org/article/077b54c4c6944878a6711e8313f6856c |
| رقم الأكسشن: | edsdoj.077b54c4c6944878a6711e8313f6856c |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 14732262 |
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| DOI: | 10.22203/eCM.v031a13 |