RT Journal Article
SR Electronic
T1 Controlling Cell-Free Gene Expression Behavior by Tuning Membrane Transport Properties
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 604454
DO 10.1101/604454
A1 Patrick M. Caveney
A1 Rosemary M. Dabbs
A1 William T. McClintic
A1 S. Elizabeth Norred
A1 C. Patrick Collier
A1 Michael L. Simpson
YR 2019
UL http://biorxiv.org/content/early/2019/04/10/604454.abstract
AB Controlled transport of molecules across boundaries for energy exchange, sensing, and communication is an essential step toward cell-like synthetic systems. This communication between the gene expression compartment and the external environment requires reaction chambers that are permeable to molecular species that influence expression. In lipid vesicle reaction chambers, species that support expression – from small ions to amino acids – may diffuse across membranes and amplify protein production. However, vesicle-to-vesicle variation in membrane permeability may lead to low total expression and high variability in this expression. We demonstrate a simple optical treatment method that greatly reduces the variability in membrane permeability. When transport across the membrane was essential for expression, this optical treatment increased mean expression level by ~6-fold and reduced expression variability by nearly two orders of magnitude. These results demonstrate membrane engineering may enable essential steps toward cell-like synthetic systems. The experimental platform described here provides a means of understanding controlled transport motifs in individual cells and groups of cells working cooperatively through cell-to-cell molecular signaling.