PUBLICATION
ORMOPLEXEs for gene therapy: In vitro and in vivo assays
- Authors
- Matos, J.C., Soares, A.R., Domingues, I., Monteiro, G.A., Gonçalves, M.C.
- ID
- ZDB-PUB-160405-6
- Date
- 2016
- Source
- Materials science & engineering. C, Materials for biological applications 63: 546-53 (Journal)
- Registered Authors
- Soares, Amadeu
- Keywords
- Gene therapy, Nanoparticles, Non-viral vector, ORMOPLEXE, ORMOSIL, Sol-gel, Transfection
- MeSH Terms
-
- Amines/chemistry
- Animals
- CHO Cells
- Cricetinae
- Cricetulus
- Electrophoretic Mobility Shift Assay
- Embryo, Nonmammalian/drug effects
- Embryo, Nonmammalian/metabolism
- Gene Transfer Techniques*
- Microscopy, Confocal
- Microscopy, Electron, Transmission
- Nanoparticles/chemistry*
- Nanoparticles/toxicity
- Nanoparticles/ultrastructure
- Particle Size
- Plasmids/genetics
- Plasmids/metabolism
- Silicon Dioxide/chemistry*
- Spectroscopy, Fourier Transform Infrared
- Toxicity Tests
- Zebrafish/growth & development
- PubMed
- 27040249 Full text @ Mater Sci Eng C Mater Biol Appl
Citation
Matos, J.C., Soares, A.R., Domingues, I., Monteiro, G.A., Gonçalves, M.C. (2016) ORMOPLEXEs for gene therapy: In vitro and in vivo assays. Materials science & engineering. C, Materials for biological applications. 63:546-53.
Abstract
Gene therapy stays on the cutting edge of biomedical research, being the design of the optimal gene delivery vector one of the key requests. Silica-based nanoparticles (NPs) have emerged as promising non-viral gene delivery vector, due to their high biocompatibility, nontoxicity, non-immunogenicity, biodegradability and enormous bioconjugation versatility. In this work a sol-gel methodology for the synthesis of amino-functionalized silica NPs (NH2-ORMOSIL NPs) was optimized, and NPs were characterized by TEM and FTIR. In a first step NH2-ORMOSIL NPs were bioconjugated with a plasmid DNA, pVAX1-GFP, assembling an ORMOPLEXE, confirmed by agarose gel electrophoresis. In a second step, in vitro studies have been performed with cultured CHO cells, where ORMOPLEXEs transfection was proved by CLSM. In vivo transfection efficiency and bio-distribution were performed in Zebrafish (Danio rerio) embryos, assessed by FM. Finally, NPs ecotoxicity was studied in zebrafish embryos by following the mortality and developmental endpoints.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping