Grant: Discovery Grants Program - Individual

This research program aims to advance analytical technology for studying DNA mutations and molecular interactions. The proposed research focuses on: (1) developing analytical technologies and ultrasensitive assays for specific DNA mutations; (2) developing techniques for imaging DNA mutations and molecular interactions; and (3) applying the new assays and techniques to the detection and imaging of target molecules in live cells. Analytical techniques for measuring DNA mutations are essential for examining the development and progression of genetic diseases, elucidating oncogenes and cancer suppressor genes, identifying drug-target or disease-resistance genes, and correcting disease-causing mutations. The first objective of this research is to develop new technologies and assays for measuring specific DNA mutations. This technology uses two rationally designed molecular probes, one binding to the DNA damage and the other binding to the specific gene. Binding of both probes to the same DNA molecule produces a new detectable DNA assembly. The ultra-sensitive technique will be applied to the detection of specific DNA damage caused by environmental and clinical agents, such as ultraviolet light, environmental contaminants, and cancer treatment drugs. The second objective is to establish a new method for imaging DNA mutations in live cells. The new method takes advantage of a powerful genome engineering technology, called CRISPR. This technology uses a set of molecules to locate the target site in the genome, make cuts to the DNA at the target site, and then edit and repair the cuts, accomplishing genome editing at the target site. In this research, the unique ability of CRISPR to locate and bind to specific sites of the genome will be used to deliver a specially designed fluorescence probe to the target site. Binding of this probe to the specific mutations in the DNA will turn on the fluorescence. Monitoring of the fluorescence generated in the cells will enable real-time imaging of the mutations. The third objective is to extend the applications of the new techniques to studying molecular interactions and visualizing molecular processes in the cells, such as how viruses are replicated. The concept and strategy of the proposed analytical technologies and assays are also applicable to many other groups of compounds of biochemical, clinical, and environmental importance. This research deals with the development of bioanalytical technologies that will significantly impact research on cancer, molecular biology, and toxicology. The proposed research will provide excellent training opportunities for students and postdoctoral fellows of diverse background, providing them with an inclusive environment and opportunities to learn and to contribute to the development of state-of-the-art analytical technologies which are in demand due to their immediate applications and future potential.