Gene Regulation,Stem Cells&Cancer Progr. Boxuan Zhao

02/07/202612:30PRBBGene Regulation,Stem Cells&Cancer Progr.Boxuan ZhaoUniversity of IllinoisMapping the Communicatome: RNA-Barcoding Technologies to Decode Cell-Cell Connections at Single-Cell ResolutionHost: Tagliaferro, BarbaraAbstract:How cells communicate, whether through synapses, contact-dependent signals, or secreted factors, underlies diverse biological processes ranging from neural circuit function to tumor progression. Yet for many cell-cell connections, the identity of the partners involved and the molecular content of those interactions remain poorly characterized, in part because tools for systematically resolving connectivity at the level of single cells have been limited. We are developing a suite of sequencing-based technologies to help close this gap. Connectome-seq maps synaptic connectivity at single-cell resolution by combining split-fluorescent protein reconstitution across synaptic clefts with RNA barcoding, allowing simultaneous identification of connected pre- and postsynaptic neurons from complex tissue. Communicatome-seq extends this framework to heterotypic cell-cell contacts, including neuron-glia and neuron-cancer interactions, using modular contact sensors that reconstitute selectively at physical interfaces. Complementing these connectivity maps, APEX2-based proximity labeling profiles the local proteome and transcriptome at synaptic microdomains, providing molecular characterization of specific connection sites. We are also developing RNA linking methods to join two RNAs into a single sequence-preserving cDNA, extending the RNA-barcoding readout behind these platforms toward mapping RNA and protein interactions directly. Together, these complementary approaches aim to advance the systematic mapping of the communicatome in the central nervous system and other tissues.