Developing a High-Throughput Single-Cell Screening Platform to Optimize Spatial and Temporal Properties of Mammalian Synthetic Circuits and Parts

Biological entities actively move in space and time. Therefore, it is imperative to optimize spatial and temporal properties such as cell-to-cell interaction and gene expression kinetics etc. to successfully repurpose synthetic genetic circuits and parts as therapeutics and diagnostics. Directed evolution is commonly used to screen libraries of variants for desirable properties. However, screening for complex spatial and temporal properties at high-throughput is not available and thereby make the optimization of these properties slow and tedious. To address this problem, we developed SPOTlight, a versatile high-throughput technique to isolate individual mammalian cells with unique spatiotemporal profiles from heterogeneous populations. SPOTlight relies on imaging visual phenotypes by microscopy, precise optical tagging of single target cells, and retrieval of tagged cells by fluorescence-activated cell sorting. We anticipate that the versatility of SPOTlight will facilitate its deployment to rapidly screen for synthetic circuits and parts with optimized spatial and temporal properties.