In Vivo Janelia Fluor dye bioavailability
The development and utility of chemigenetic sensors (eg. Voltron, HaloCaMP, WHaloCaMP) is dependent upon the reliable delivery of HaloTag ligand- or SNAP tag ligand-dyes to an in vivo tissue target. We are characterizing the bioavailability of the current Janelia Fluor dyes and are using that data to further guide the design of additional fluorescent dyes.
Calcium sensor applications
The GCaMP series of calcium sensors have a legacy at Janelia. These calcium sensors were developed as proxies for monitoring neuronal activity. We have taken our institutional knowledge of these sensors and adapted their use in alternative applications to monitor calcium fluctuations in organelles, in the extracellular matrix, and as cell death indicators.
Metabolic and signaling sensor development
We continue to use our expertise in protein engineering to improve existing metabolic and signaling sensors and to demonstrate their utility in a variety of experimental systems. This includes recent development of fluorescent sensors for glucose, ATP, cAMP and aspartate.
Membrane protein expression
The efficient expression and targeting of engineered proteins to the plasma membrane of cells is often challenging. We seek to develop strategies to improve membrane expression efficiency through sequence elements that enhance translation and processing through the ER and Golgi trafficking pathways.
Characterization of novel fluorescent dyes in experimental systems
We work closely with the Lavis lab to validate novel dyes and their biological application. This includes both dye fluorescence properties as well as specificity ligands which target the dyes to places of interest.