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Title: Biological applications of DNA cages
Author: Walsh, Anthony S.
ISNI:       0000 0004 2721 4607
Awarding Body: Oxford University
Current Institution: University of Oxford
Date of Award: 2011
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DNA cages are nanometre-scale polyhedral structures formed by self-assembly from synthetic DNA oligonucleotides. In this thesis we describe the development of DNA cages for biological applications, with a particular focus on their potential as agents for the delivery and control of macromolecular cargoes within cells. We report an investigation of the ability of a model cage, a DNA tetrahedron, to enter live cultured mammalian cells. Substantial uptake of tetrahedra into cells was observed both when the cells were treated with tetrahedra alone and when the cells were treated with a mixture of tetrahedra and a transfection reagent. Analysis of the subcellular localization of transfected tetrahedra using con focal microscopy and organelle staining indicates that the cages are located in the cytoplasm. FRET experiments indicate that the DNA cages remain substantially intact within the cells for at least 48 hours after transfection. We describe an investigation into the ability of DNA cages to carry macromolecular cargoes (proteins and functional oligonucleotides) into mammalian cells. Delivery of fluorescently labelled protein into cells was achieved with the aid of DNA cages (DNA octahedra). DNA cages with structures capable of opening in response to specific molecular signals (including full length mRNA molecules) were developed. We also demonstrated the ability of DNA cages to functionally encapsulate single enzyme molecules within their interior cavities. It is proposed that these studies provide a promising proof of principle of the potential of DNA cages as agents for the delivery and control of macromolecular cargoes within cells. Potential applications of such devices include in vivo imaging and the targeted delivery of therapeutic macromolecules into living cells.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available