This proposal will explore the use of single-molecule fluorescence spectroscopy to study the solution structure of small single-stranded and double-stranded RNAs. The long-term funding enabled by the research proposed here will provide new instrumentation to Dr. Krueger, allowing him to continue this work in the future and to incorporate the research ideas into the teaching curriculum at Hope College, providing lasting benefit to Hope College and its students. This is potentially high impact research that may realize tangible benefit to human health through improved understanding of regulation through RNA. The use of single-molecule fluorescence permits the structural diversity and dynamics of the RNAs to be understood in a way that is complementary to bulk time-resolved studies that will also be executed in Dr. Krueger's laboratory. The Krueger group will also use molecular dynamics simulations to model the RNA structural dynamics helping to connect the two spectroscopic techniques and to provide more detail. GLCA New Directions Initiative funding will allow Dr. Krueger to carry the work all the way from acquisition of samples to first publication, providing the foundation necessary for him to seek long-term funding from government sources.