This project will design and apply new probes to detect neurodegenerative diseases caused by the expansion of short sequences in human gene. The neurodegenerative diseases (e.g. Friedreich Ataxia, Huntington's Disease, Amyotrophic Lateral Sclerosis) caused by such expansions are heritable and lead to death at early age. Current methods are limited to detecting small fragments along with imprecise interpretation of the actual length of the suspected sequence. In the proposed research, the principal investigator will engineer short nucleic acid probes to detect long neurodegenerative DNA sequence targets with high dynamic range and resolution. This research project will span the fields of chemistry, biology, and engineering and provide a highly interdisciplinary training experience to high school, undergraduate, and graduate students. This project will also contribute to public scientific literacy through workshops and Science Cafe discussion at Carbondale Science Center.
In human gene, repeat sequence expansion occurring in tandem cause more than two dozen neurodegenerative and neuromuscular diseases such as Friedreich Ataxia, Huntington's Disease, Amyotrophic Lateral Sclerosis etc. These diseases are genetically transferrable and leading to death at early age. Current methods lack direct and precise interpretation of repeat lengths in a suspected gene. The principal investigator will direct an experimental research project to develop novel oligonucleotide probes for the impedance based detection of pathological lengths of neurodegenerative repeats (e.g. GAA, CAG, GGGGCC repeats) in order to enhance dynamic range, sensitivity, and resolution of detection. The central hypothesis for this project is that oligonucleotide probes with non-tandem complementary repeats and rigid backbones can capture 10x larger complementary targets and transduce into an impedance signal, which can be proportionally correlated with the number of repeats in pathogenic range. The principal investigator will a) design probes to correlate impedance signal with pathogenic length of repeat sequences, b) test detection range of the probes for neurodegenerative repeats, c) test specificity of the designed probes by challenging them with non-specific repeat sequences, and d) detect neurodegenerative repeats in real samples. The challenge in the project is to achieve highly linear detection range up to pathogenic length with single repeat resolution due to complex secondary structures of long repeat targets, which makes this proposal high risk and high payoff. The broader impact of this proposal is multidimensional. First, this fundamental research will provide a detection tool to improve public health and biomedical research. Second, it involves integration of major STEM fields including chemistry, biology, and engineering. Third, it will train STEM workforce for nucleic acid biosensing research at high school, undergraduate, and graduate level. Fourth, general public will be educated by discussing research output in Science Cafe at The Carbondale Science Center.
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.
