CAREER: Structural Implications of Nucleic Acid Nanoparticles on their Transcription, Translation, and Transport

About This Grant

Delivering therapeutic drug molecules or genes to cells requires nanoscale carriers that can interact with proteins inside the cell and navigate to the nucleus, if needed. Nanoparticles made entirely out of synthetic deoxyribonucleic acid strands have proven useful in building such delivery systems. Although they work well for drug delivery, their behavior inside the cell is not fully understood, which presents challenges to improving nanoparticles for specific applications. This project will encode synthetic deoxyribonucleic acid nanoparticles with gene sequences and measure the extent to which these nanoparticles interact with the cell’s protein building machinery to make a protein of choice. Various bioengineering and fluorescence tools will be used to study the nanoparticles in living cells. Effects of shape, design, and other features of the nanoparticles on their behavior and function as gene delivery systems will be determined. The team will engage local high school students in research for 10-week long programs at Case Western Reserve University. The project will also support educational module design using mixed reality technology to teach topics in bioconjugate chemistry. Nanoparticles created from self-assembly of synthetic deoxyribonucleic acids (referred to as DNA origami nanoparticles) are being developed for targeted delivery of drugs, bioimaging probes, and functional nucleic acids. However, the stability of these nanoparticles and how they interface with cellular processes are not understood. The main limitation thus far has been a lack of bioanalytical tools that provide sufficient resolution to study nucleic acid nanoparticles in physiological conditions. Without understanding the mechanism of nanoparticle design and its intracellular functionality, engineering application-specific nanoparticles is not feasible. The goal of this project is to determine the role of nanoparticle shape and sequence layout in their ability to express proteins in cells. The mechanistic hypothesis is that predictable shape and sequence layout of gene-encoded nanoparticles will sterically tune transcription processivity and ultimately affect their stability and function. The research will measure how gene-encoded nucleic acid nanoparticle processing depends on their shape, gene sequence routing within the nanoparticle, and their transport across the nuclear membrane. An integrated educational plan will have three synergistic activities, namely, (1) high school student workshop on how to manifest a DNA nanoparticle using makerspaces, (2) 3-dimensional printable models to illustrate how fluorescent molecules interact on DNA nanoparticles, and (3) a mixed reality teaching module to transform bioconjugate chemistry education. The project will catalyze a long-term research program that trains the next generation of scientists in bioengineering, nanotechnology, and analytical chemistry. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.