High-Performance Liquid Chromatography for DNA Structure Purification and Characterization

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We urgently request a high-performance liquid chromatography (HPLC) instrument to replace our current HPLC system, which is 9 years old and has recently broken down. This has significantly slowed down the research of 20 HQP in my research group, as all the DNA conjugates that we synthesize require HPLC purification and characterization. The requested instrument will support our research program which focuses on developing DNA structures for biological applications. Specifically, this proposal introduces a new, highly scalable method to develop stimuli-responsive DNA hydrogels for tissue regeneration, antimicrobial and biosensing applications. We recently discovered that a small, inexpensive and non-toxic molecule, cyanuric acid, can induce DNA to assemble into a triple helix that grows into long nanofibers. We propose to chemically polymerize adenosine to form poly(adenine) DNA at scale, followed by the addition of cyanuric acid derivatives and crosslinking, to produce biocompatible hydrogels in a single pot with environmentally benign reagents and solvents. We will develop these hydrogels to promote cellular growth, and to respond to multiple biological stimuli through changes in mechanical and drug release properties. In developing the first small molecule-promoted DNA hydrogel, we expand the application space of biocompatible materials: our hydrogels are scalable, mechanically robust, stimuli-responsive, and recyclable, enabling a new, green methodology for the development of DNA hydrogel materials for widespread medical use. Our second project involves the development of sequence-controlled polymers as carriers of nucleic acid therapeutics. We aim to overcome important barriers that prevent their translation to the clinic, such as poor cellular uptake, and entrapment in organelles called endosomes. DNA strands conjugated to sequence-controlled polymers assemble in aqueous media to form monodisperse spherical structures, with a hydrophobic core and a dense nucleic acid corona (spherical nucleic acids, SNAs). We propose to develop these structures are highly effective delivery vehicles, where their nucleic acid exterior silences the expression of disease-causing proteins even in naked form, and can efficiently escape endosomal entrapment. We will also perform high-throughput cell selection studies, using a new class of DNA-encoded polymers developed in our lab. The objective is to select polymers that mediate the escape of nucleic acid therapies from endosomes, and that allow them to operate in the cell cytosol. Overall, the requested HPLC will ensure that productivity in our laboratory is not slowed down, and it will accelerate the discovery of new DNA biomaterials for tissue regeneration and for gene therapy. It will help train HQPs in nucleic acid research, an area that is critically important to ensure Canada's preparedness to any future pandemic.