Study of DNA-cationic polymer brush interactions for gene delivery
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1
Queen Mary University of London, Institute of Bioengineering and School of Engineering and Materials Science, United Kingdom
Gene delivery relies on the encapsulation of DNA coding for a gene of interest, which is ideally delivered to target cells to provide the treatment of both inherited and acquired diseases [1]. Particular interest has focused in recent years on the development of efficient nonviral vectors, which are less hazardous in terms of antigen-specific immune responses compared to viral vectors [2]. Encapsulation of naked DNA by cationic polymers such as poly (2-dimethylaminoethyl methacrylate) (PDMAEMA) which is positively charged at physiological pH, has been extensively investigated for its potential as a gene delivery vector. Among different macromolecular architectures of PDMAEMA based cationic polymers (e.g. block, star or graft), brush structures have been recognised as particularly promising systems as they allow the simultaneous control of polymer architecture, vector size and shape, surface chemistry and offer the ability of live monitoring [3].
This project focuses on the preparation of PDMAEMA based polymer brushes on flat surfaces and silica particles to understand the behaviour in different buffer conditions, their interaction with DNA molecules as well as the transfection efficiency of the SiO2-PDMAEMA/DNA polyplex. Firstly, PDMAEMA brushes with different thicknesses and grafting densities were synthesised via ATRP on silicon wafer and gold substrates to study the brush behaviours (e.g. swelling and collapse via ellipsometry) under different buffers conditions used for the formation of DNA- polyplexes. We used surface plasmon resonance (SPR) and in situ ellipsometry to further investigate how DNA interacts with PDMAEMA brushes under those conditions. We found that DNA binding strongly depends on buffer conditions,brush thickness and density and correlates with transfection efficiency with epidermal cells.
In addition, to track the fate of particle-DNA complexes during the DNA delivery and transfection process, we developed a method combining layer-by-layer self-assembly with ATRP, allowing us to label PDMAEMA brush decorated silica particles with fluorescent tags without changing any chemical or structural properties of the brush-particles. This allowed us to study simultaneously the fate of brush-particles and DNA molecules in situ during cell transfection experiments.
China Scholarship Council
References:
[1] Conwell, C. C. & Huang, L. Recent advances in non-viral gene delivery. Adv Genet 53, 3-18 (2005).
[2] Kataoka, K. & Harashima, H. Gene delivery systems: viral vs. non-viral vectors. Adv Drug Deliver Rev 52, 151-151 (2001).
[3] Krishnamoorthy, M., Hakobyan, S., Ramstedt, M. & Gautrot, J. E. Surface-Initiated Polymer Brushes in the Biomedical Field: Applications in Membrane Science, Biosensing, Cell Culture, Regenerative Medicine and Antibacterial Coatings. Chem Rev 114, 10976-11026 (2014).
Keywords:
DNA,
Surface modification,
gene delivery,
polymer brush
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
Poster
Topic:
Biomaterials in gene therapy
Citation:
Li
D,
Krishnamoorthy
M and
Gautrot
J
(2016). Study of DNA-cationic polymer brush interactions for gene delivery.
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.01765
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.
*
Correspondence:
Dr. Danyang Li, Queen Mary University of London, Institute of Bioengineering and School of Engineering and Materials Science, London, United Kingdom, danyang.li@qmul.ac.uk
Dr. Mahentha Krishnamoorthy, Queen Mary University of London, Institute of Bioengineering and School of Engineering and Materials Science, London, United Kingdom, m.krishnamoorthy@qmul.ac.uk
Dr. Julien Gautrot, Queen Mary University of London, Institute of Bioengineering and School of Engineering and Materials Science, London, United Kingdom, j.gautrot@qmul.ac.uk