Tucker Carrocci
Title: New Methods for Fluorescently Labeling RNA In Vitro and In Vivo
Advisor: Aaron Hoskins
Program: Integrated Program in Biochemistry (IPiB)
Fluorescence microscopy has proven incredibly useful for understanding biological
systems. However, the inability to easily label RNAs with fluorophores has limited this
technique in studies of RNA biology. We are developing new tools for labeling RNAs
with fluorophores in vivo and in vitro. Using fusions of the phage MS2 protein and
Escherichia coli dihydrofolate reductase, we can now label RNAs in living yeast cells
with fluorescent derivatives of trimethoprim. By exploiting protein fragment
complementation, we have further optimized the use of DHFR for RNA labeling by
reducing background fluorescence. In the test tube, we are employing deoxyribozymes to
site-specifically modify RNAs with fluorophores. This method should be applicable for
preparing long, highly modified fluorescent RNAs for biophysical studies.