Sensing and Delivery of Biomolecules with Spherical Nucleic Acid Nanoparticle Conjugates

Larkin, Isaac Nathaniel

doi:https://doi.org/10.21985/n2-sfc1-r876

Work

Sensing and Delivery of Biomolecules with Spherical Nucleic Acid Nanoparticle Conjugates

Public

Download PDF

Download All Files (.zip)

Over the past fifty years, techniques for synthesizing and manipulating matter on the 1-100 nanometer scale have led to the development of nanoparticle-based approaches to both disease diagnosis and treatment. The modification of nanoparticles with biological macromolecules such as proteins and nucleic acids has led to the development of highly sensitive detectors of disease biomarkers and has facilitated the delivery of therapeutic molecules into target tissues and cells. Challenges and opportunities in the development of biomolecule-functionalized nanoparticles include: (1) how to balance the sensitivity of a biomarker diagnostic assay against the cost and complexity of the equipment required to perform the assay; (2) developing design rules for the construction of nucleic-acid-based biosensors of small molecules; and (3) cytosolic delivery of extracellular enzymes by avoiding endosomal entrapment. Present in this thesis is an exploration of biomolecule-functionalized nanoparticles for addressing these challenges. Chapter one surveys the origins and applications of a wide variety of bio-functionalized nanoparticles. In particular, I focus on spherical nucleic acids (SNAs), nanoparticles densely functionalized with a highly oriented oligonucleotide shell, which possess structural and biological properties distinct from linear nucleic acids that have enabled the development of biosensors and new therapeutics. Chapter two details the use of antibody-functionalized gold nanoparticles to develop a dual readout assay for device-free and highly sensitive detection of an anthrax biomarker. Chapter three explores the properties and design rules of aptamer NanoFlares, a class of SNA functionalized with DNA aptamers and hybridized with small fluorophore-modified oligonucleotides, in the context of work aimed at developing an assay for small molecule biomarkers of human stress. Chapter four investigates the possibilities and challenges of adapting SNAs for the delivery of gene-editing enzymes, with the goal of using gene editing to better understand the endosomal escape of SNAs. Finally, chapter five discusses the challenges encountered during these projects and provides perspective on how researchers could build on this work going forward.

Creator