RT Journal Article
SR Electronic
T1 Molecular Transport across Lipid Membranes Controls Cell-Free Expression Level and Dynamics
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 606863
DO 10.1101/606863
A1 Patrick M. Caveney
A1 Rosemary M. Dabbs
A1 William T. McClintic
A1 C. Patrick Collier
A1 Michael L. Simpson
YR 2019
UL http://biorxiv.org/content/early/2019/04/12/606863.abstract
AB Essential steps toward synthetic cell-like systems require controlled transport of molecular species across the boundary between encapsulated expression and the external environment. When molecular species (e.g. small ions, amino acids) required for expression (i.e. expression resources) may cross this boundary, this transport process plays an important role in gene expression dynamics and expression variability. Here we show how the location (encapsulated or external) of the expression resources controls the level and the dynamics of cell-free protein expression confined in permeable lipid vesicles. Regardless of the concentration of encapsulated resources, external resources were essential for protein production. Compared to resource poor external environments, plentiful external resources increased expression by ~7-fold, and rescued expression when internal resources were lacking. Intriguingly, the location of resources and the membrane transport properties dictated expression dynamics in a manner well predicted by a simple transport-expression model. These results suggest membrane engineering as a means for spatio-temporal control of gene expression in cell-free synthetic biology applications and demonstrate a flexible experimental platform to understand the interplay between membrane transport and expression in cellular systems.