Synthetic Gene Circuits Controlling Precise Biological Pattern Formation in Multicellular-Mammalian Systems

Small molecules offer a diverse range of tools for probing fundamental principles of biological organization and hierarchy in biological physics. Yet, despite their strengths for controlling temporal biological functions, they are limited in exploring questions relevant to biological pattern formation. To address this need for studying spatiotemporal phenotypes in multicellular systems, we have created a toolbox of multiple light-inducible gene circuits that can tune gene expression and control levels of transcriptional noise at the single cell level. We accomplish this by engineering the tetracycline gene-expression system to convert light stimuli information into cellular response proteins in a spatiotemporal context with single-cell resolution. These circuits are built using the light responsive proteins LOV2 & VVD, a small peptide synthesized by the cell, and a light-inducible degradation tag. The resulting tools provide a platform for robust gene expression control in a spatiotemporal fashion, allowing easy exchange for genes of interest and probing physical biology questions in the context of cell-to-cell communication.