I have the expertise, leadership, training and motivation necessary to successfully carry out the proposed research project. I have a broad background in physics, chemistry, molecular and cell biology. As a postdoctoral fellow and then a staff scientist in Japan (1994-2002), I expressed the first GFP and BFP variants and later DsRed in mammalian cells and transgenic Drosophila flies. As an assistant professor in Colorado (2002-2005), I developed a 3-chromophore Förster resonance energy transfer (FRET) in mammalian cells, engineered photoactivatable PAmRFP1, and co-developed PSCFP and Dendra. My laboratory at Einstein (2006-present) has engineered numerous FPs, including the brightest monomeric blue TagBFPs; monomeric Fluorescent Timers; photoactivatable PAmCherry, PATagRFP and PAmKate for super-resolution PALM; large Stokes shift monomeric red (LSSmKate) and orange (LSSmOrange) FPs for 2P intravital imaging; far-red monomeric TagRFP657 and TagRFP675; and several types of near-infrared iRFP and miRFP proteins for deep-tissue imaging. I engineered FRET biosensors based on the TagBFP-TagGFP2 and LSSmOrange-mKate2 pairs, as well as split FPs and miRFPs reporters using a bimolecular fluorescence complementation (BiFC). I applied the new fiber lasers and supercontinuum white lasers to advance flow cytometry, and co-developed 2P and photoacoustic techniques for iRFP imaging. Recently, I have developed the first several near-infrared optogenetic systems for gene transcription regulation, enzymatic activity, and protein interactions.
FPs, biosensors and optogenetic constructs developed in my laboratory have been requested by ~1000 labs worldwide (~6000 plasmids provided through Addgene and ~500 directly from our laboratory). I was invited to present lectures at >50 research conferences and workshops, and >50 seminars in other institutions. I served multiple times as a reviewer for ~20 scholarly journals, including Nature and Cell families of journals. I supervised or co-supervised 12 MS and PhD students, 20 postdocs, and 5 technicians. In Einstein, I served on several committees and involved in semi-annual graduate course “Quantitative imaging of cells”. PhD students and postdocs from my laboratory regularly attend ASCB, ACS and BPS meetings. I have co-founded Fluorescent proteins resource center at Einstein through which we provide researchers with plasmids and consultations on FPs, biosensors, FP fusion constructs and imaging experiments. I am a member of Gruss-Lipper Biophotonics Center at Einstein where we build various custom imaging and flow cytometry setups, which are also available to all researchers. I collaborate or collaborated with ~10 laboratories in Einstein and with ~25 laboratories in other institutions. I was interviewed on my research or my research was featured in ~40 articles in scholarly journals. I actively participated in 5 awarded to Einstein shared instrumentation grants, including OD018218 on which I was a PI.