Electric-Field Manipulation of a Compartmentalized Cell-Free Gene Expression Reaction

ACS Synth Biol. 2018 Aug 17;7(8):1829-1833. doi: 10.1021/acssynbio.8b00160. Epub 2018 Jul 31.

Abstract

Direct electric-field manipulation of gene expression reactions would simplify the design of biochemical networks by replacing complex biomolecular interactions with push-button operations. Here, we applied a localized electric field gradient at megahertz frequency to manipulate a cell-free gene-expression reaction in a DNA compartment on a chip. We broke the spatial symmetry of a homogeneous reaction in the compartment by creating a trap for macromolecules in a region of maximal field intensity localized 50 μm from immobilized DNA. Free of biochemical regulation, we demonstrated protein synthesis oscillations by on/off switching of the electric field. In response to the field, ribosomes, RNA polymerases, and nascent RNA and proteins accumulated in the trap, and were then depleted from the DNA region where gene expression occurred. The resulting reduction in the rate of protein synthesis recovered back to steady-state when the field was off. The combination of electric field with compartmentalized cell-free gene expression reactions creates a simple, label-free approach for controlling biomolecules in space and time, opening possibilities for hybrid biological systems with a bioelectronic interface based on minimal biological parts design.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA / metabolism
  • DNA-Directed RNA Polymerases / genetics
  • DNA-Directed RNA Polymerases / metabolism
  • Electrochemical Techniques / methods*
  • Gene Expression
  • Oligonucleotide Array Sequence Analysis
  • RNA / metabolism
  • Ribosomes / metabolism

Substances

  • RNA
  • DNA
  • DNA-Directed RNA Polymerases