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DNA Origami as a Technique to Build Virus Engulfing Shell Complexes

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dc.contributor.advisor Veneziano, Remi
dc.contributor.author Holland, William Alexander
dc.creator Holland, William Alexander
dc.date 2022-01-21
dc.date.accessioned 2022-03-29T01:45:47Z
dc.date.available 2022-03-29T01:45:47Z
dc.identifier.uri http://hdl.handle.net/1920/12773
dc.description.abstract Deoxyribonucleic acid (DNA) is the molecule used by all living organisms to carry their genetic information and pass on their heritable traits between generations. DNA molecules have also other intrinsic properties that make them promising building blocks to assemble novel bio-nanomaterial. Within the realm of nanotechnology exists a subfield known as structural DNA nanotechnology that uses the DNA molecule as a building block to precisely assemble nanostructures with prescribed features. DNA origami is one of the most versatile techniques developed in the DNA nanotechnology field. Indeed, DNA origami is an assembly technique, which utilizes a single stranded DNA scaffold that can be hybridized by many short oligonucleotides called staple strands to fold the nanoparticle into any desired shape in the 10 to 100 nm scale. These nanoparticles can be made into any shapes including 2D arrays and 3D nanoparticles that can be used for drug delivery, vaccine development, and viral capture among other applications. In addition, DNA is also easily functionalized with organic and inorganic molecules such as fluorophores and targeting moieties, which can be useful in visualization or tracking. The focus of this research is to leverage the advantages of DNA origami to design and assemble nanoparticles capable of enveloping viruses based on their respective capsid’s geometry and glycoprotein organization as a novel viral therapy technique. In this investigation, the rabies virus is used as a model of target. If the structural parameters of the array and the organization of the capturing moieties match with those of the viral capsid, then the DNA array nanoparticles should possess the capacity to efficiently bind to the Rabies Virus. Here, I designed and synthetized multiple nanoscale designs that meet the viral parameters of the rabies virus using the Tiamat software. Agarose gel electrophoresis was used to validate proper folding of the DNA origami nanoparticles. In a second time, gold beads modified with streptavidin was used to mimic the virus and serve as proof of principle with biotinylated DNA nanoarrays. The binding was validated with dynamic light scattering (DLS) and zeta potential measurements that show increase in size and change in the surface charge matching with coverage with DNA. All of these tests indicated that the origami nanoparticle had successfully bound to the gold-streptavidin complex and is ready to be used with viral capsids. en_US
dc.language.iso en en_US
dc.subject DNA en_US
dc.subject shell en_US
dc.subject virus en_US
dc.subject DNA Origami en_US
dc.subject engulfing en_US
dc.subject rabies en_US
dc.title DNA Origami as a Technique to Build Virus Engulfing Shell Complexes en_US
dc.type Thesis en_US
thesis.degree.name Master of Science in Bioengineering en_US
thesis.degree.level Master's en_US
thesis.degree.discipline Bioengineering en_US
thesis.degree.grantor George Mason University en_US


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