Crowding-Induced Spatial Organization of Gene Expression in Cell-Sized Vesicles

• DOI: 10.1021/acssynbio.2c00336
• Publication Date: 2022-10-19T20:05:12Z
• Authors: Gaurav Chauhan; S. Elizabeth Norred; Rosemary M. Dabbs; Patrick M. Caveney; John K. Vincent G...; C. Patrick Collier; Michael L. Simpson; Steven M. Abel
• Keywords: Macromolecular Crowding; Crowding

Cell-free protein synthesis is an important tool for studying gene expression and harnessing it applications. In cells, regulated in part by the spatial organization of transcription translation. Unfortunately, current cell-free approaches are unable to control molecular components needed expression, which limits ability probe utilize its effects. Here, we show, using complementary computational experimental approaches, that macromolecular crowding can be used translational efficiency cell-sized vesicles. Computer simulations imaging experiments reveal that, as increased, DNA plasmids become localized at inner surface Ribosomes, contrast, remain uniformly distributed, demonstrating differentially organize expression. We further carried out reactions vesicles quantified mRNA abundance. At sufficiently high levels crowding, observed localization near vesicle surfaces, a decrease abundance, anomalous scaling abundance function size. These results consistent with causing altered slower diffusion. Our work demonstrates straightforward way provides insight into regulation cells.