title: Molecular crowding shapes gene expression in synthetic cellular nanosystems.
abstract: The integration of synthetic and cell-free biology has made tremendous strides towards creating artificial cellular nanosystems using concepts from solution-based chemistry, where only the concentrations of reacting species modulate gene expression rates. However, it is known that macromolecular crowding, a key feature in natural cells, can dramatically influence biochemical kinetics via volume exclusion effects, which reduce diffusion rates and enhance binding rates of macromolecules. Here, we demonstrate that macromolecular crowding can increase the robustness of gene expression by integrating synthetic cellular components of biological circuits and artificial cellular nanosystems. Furthermore, we reveal how ubiquitous cellular modules, including genetic components, a negative feedback loop and the size of the crowding molecules can fine-tune gene circuit response to molecular crowding. By bridging a key gap between artificial and living cells, our work has implications for efficient and robust control of both synthetic and natural cellular circuits.
authors.affiliation: Lane Center for Computational Biology, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213, USA.
authors.forename: Cheemeng
authors.initials: C
authors.lastname: Tan
authors.forename: Saumya
authors.initials: S
authors.lastname: Saurabh
authors.forename: Marcel P
authors.initials: MP
authors.lastname: Bruchez
authors.forename: Russell
authors.initials: R
authors.lastname: Schwartz
authors.forename: Philip
authors.initials: P
authors.lastname: Leduc
mesh_terms: D017123:Bacteriophage T7 / Q000235:genetics / Q000378:metabolism; D003198:Computer Simulation; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000302:isolation & purification*; D004058:Diffusion; D015870:Gene Expression; D046911:Macromolecular Substances / Q000737:chemistry; D015394:Molecular Structure; D036103:Nanotechnology / Q000379:methods*; D011401:Promoter Regions, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D046911:Macromolecular Substances; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases