Source: Dissertations Abstracts International, Volume: 69-10, Section: B
Publisher info.: Dissertation/Thesis
Advisor: Luo, Dan
Thesis (Ph.D.)--Cornell University, 2007
Includes bibliographical references
DNA is generally recognized for its functional role as a medium for storing genetic information. Recently, however, it has also been utilized as a structural scaffold for new DNA nanostructures due to its characteristic specificity, flexibility and manipulability. Here, new DNA engineered materials are reported-dendrimer-like DNA (DL-DNA), DNA hydrogel, and protein-producing DNA hydrogel (P-gel). A variety of DNA building blocks (e.g. X-, Y-, T-DNA) were used as structural motifs for the construction of these DNA materials. DL-DNA was constructed by assembling Y-DNA building blocks step by step onto a solid support through both sticky end cohesion and enzymatic ligation. With fluorophore labeling, this DL-DNA was utilized as a nanobarcode for multiplexed pathogen detection. X-, Y-, and T-DNA building blocks were assembled into three-dimensional DNA hydrogels through an enzymatic reaction. By varying the initial concentration and type of DNA building blocks, the chemical and physical properties of these DNA hydrogels could be precisely tuned. The resulting DNA hydrogels were patterned into various shapes in the micro and millimeter scale. These DNA hydrogels were explored for use as a new controlled drug delivery system, a cell culture matrix, and an in vitro protein production platform. The release of insulin drug from DNA hydrogel demonstrated a strong dependence on the type of DNA building blocks composing the hydrogel. As a cell culture matrix, DNA hydrogel was used to encapsulate CHO cells in situ. X-DNA building blocks, functioning as both crosslinkers and building blocks, were enzymatically crosslinked with genes encoding a model protein, Renilla luciferase (Rluc) to form a novel protein-producing DNA hydrogel (P-gel). This P-gel produced up to 325 μg of Rluc per μg of plasmid, a yield approximately 150 times higher than that of a conventional solution-based cell-free systems at the same plasmid concentration. The higher productivity of P-gel was also demonstrated by green fluorescence protein (AcGFP) and chloramphenicol acetyl transferase (CAT) as proof-of-concept proteins. The substantial improvement in protein production was attributed to enhanced mRNA transcription due to a higher local gene concentration, and a protecting effect caused by the networked DNA hydrogel against degradation of incorporated genes
Electronic reproduction. Ann Arbor, Mich. : ProQuest, 2020
