In Vitro Characterization and Assessment of Novel DNA Origami-Aptamer Modification Intended for Targeting Glioblastoma-Associated Biomarker Nucleolin
| dc.contributor.advisor | Veneziano, Rémi | |
| dc.contributor.author | O'Shea, Bailey Jacob | |
| dc.creator | O'Shea, Bailey Jacob | |
| dc.date | 2021-07-30 | |
| dc.date.accessioned | 2022-05-13T21:41:36Z | |
| dc.date.available | 2022-05-13T21:41:36Z | |
| dc.description.abstract | Ample amounts of literature have demonstrated the practicality of functionalizing DNA origami with aptamers—single-stranded DNA or RNA target-specific ligands with similar binding affinities to monoclonal antibodies—to effectively target and treat a wide range of malignancies. When functionalizing origami with aptamers, the current method relies on the use of staple strands that can be used to hybridize the aptamer of choice modified with a complementary sequence or modified with overhangs bearing the aptamer sequences. These approaches require additional steps during both the computational design and in vitro synthesis stage and limits our capacity of functionalization. Herein, we provide a novel technique that overcomes these additional steps while allowing for the functionalization of origami with aptamers. We demonstrate by simply designing single-stranded DNA (ssDNA) aptamer “loops”, we are able to introduce aptamers at precise locations within the scaffold sequence while retaining target binding without the need to employ modified staples. Through the use of agarose gel electrophoresis, we were able to show the characterization of two structures—a 42 bp edge 2-Dimensional triangle and 3-Dimensional tetrahedron—presenting the well-studied thrombin 29-mer and AS1411 26-mer nucleolin aptamer. After exposing thrombin to our origami-aptamer modified constructs, we observed each structure had the ability to effectively bind to thrombin. Our new sequence-aptamer functionalization technique may pave the way for simple design of readily functionalized DNA origami nanoparticles for a wide range of fields including diagnostic and therapeutic-related research. | |
| dc.identifier.uri | https://hdl.handle.net/1920/12842 | |
| dc.language.iso | en | |
| dc.subject | DNA Origami | |
| dc.subject | Aptamer | |
| dc.subject | DNA nanotechnology | |
| dc.subject | DNA Origami-aptamer | |
| dc.subject | Thrombin aptamer | |
| dc.subject | In Vitro Characterization and Assessment of Novel DNA Origami-Aptamer Modification Intended for Targeting Glioblastoma-Associated Biomarker Nucleolin | |
| dc.title | In Vitro Characterization and Assessment of Novel DNA Origami-Aptamer Modification Intended for Targeting Glioblastoma-Associated Biomarker Nucleolin | |
| dc.type | Thesis | |
| thesis.degree.discipline | Bioengineering | |
| thesis.degree.grantor | George Mason University | |
| thesis.degree.level | Master's | |
| thesis.degree.name | Master of Science in Bioengineering |