Developing Selective Conjugation Reactions for Applications in > Cellular Imaging and Synthetic Biology

Our research group is interested in the development of of in-situ bioconjugation reactions through the pursuit of 2 major research efforts: 1. The study of new bioorthogonal reactions for applications in cellular imaging, and 2. Developing synthetic cells through artificial phospholipid synthesis and modification. For cellular imaging applications, we have focused on advancing bioorthogonal tetrazine ligations with strained dienophiles. We have developed novel synthetic routes to tetrazines, leading to the synthesis of fluorogenic probes that increase over 100-times in fluorescence intensity after ligation. Metabolic imaging applications using tetrazines have been made possible by new dienophiles, such as methyl-cyclopropene mini-tags. Recently, we have introduced tetrazine-mediated transfer (TMT) reactions, which enable DNA/RNA template driven turnover of fluorogenic reactions and the detection of clinically relevant microRNAs. We also have a strong interest in applying covalent coupling reactions to the formation and modification of phospholipid membranes. We have utilized chemoselective reactions, such as copper-catalyzed triazole formation or the native chemical ligation, to drive the de novo synthesis of phospholipid membranes. To interface synthetic membranes proteins, we have recently developed protein reactive membrane anchors utilizing SNAP-tags. Light driven spatiotemporal control over proteoliposome formation is possible using photocaged membrane anchors. Continued development of selective bioconjugation reactions will facilitate the interfacing of synthetic molecules with biological systems for new capabilities.