Engineering functionalized diamond nanoparticles for intracellular delivery of nucleic acids

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The objective of my research program is to engineer novel nanomaterials that carry nucleic acids into mammalian cells. Gene delivery has been a significant research area for the past two decades, but the mainstream gene delivery nanomaterials, lipids and polymers pose limitations with respect to their stability, efficiency and toxicity. My research program focuses on the use of solid--core nanoparticles, such as nanodiamonds, to overcome these barriers. Nanodiamonds have a uniform size distribution and a large surface area available for chemical modifications. They are nontoxic in doses necessary for gene delivery. My research strategy is to chemically link amino acids to the nanodiamond in order to facilitate their interaction with nucleic acids. The novelty of this program lies in the approaches to the synthesis and characterization of nanoparticles conjugated with bioactive molecules, amino acids and elucidation of their beneficial and detrimental interactions with mammalian cells, organs and whole body. The short--term research objectives build on my previous work: design of self--assembling lipid--based and diamond--based nanoparticles. To achieve our goal, we decorate the surface of nanodiamond with amino acids resulting in functionalized nanoparticles amenable to binding nucleic acids. We will assess the ability of these functionalized nanoparticles to deliver nucleic acids into model cells. The biodistribution of various functionalized nanodiamonds will be monitored. Relationships between structure/architecture - delivery efficiency/toxicity will be established. This will lead to the selection of candidate vectors with optimal properties (improved binding properties, cellular uptake and high activity with no or minimal toxicity, and stability in biological environment) for delivering nucleic acids into cells. To study the architecture of the delivery systems, high--resolution microscopy, flow cytometry and synchrotron-based techniques will be used. Correlating the surface properties of these nanoparticles with the delivery efficiency will allow for mapping universal and carrier--specific delivery mechanisms. Long term, we will explore how the basic knowledge on chemical functionalization of the nanodiamond particles could be expanded to other solid--core and soft nanoparticles, establishing universal delivery principles for nucleic acids. Correlation of biodistribution to surface properties will be used as a model for predicting the biological fate of other man-made nanoparticles with similar surface properties. This research program will advance knowledge of the use and safety of the functionalized diamond--core nanoparticles as delivery systems for nucleic acids. Optimization of the characteristics of these delivery systems will benefit nanotechnology (a global market of $1.2B), eventually leading to gene therapy in humans and animals.