A multiplexed, electrochemical interface for gene-circuit-based sensors

Nat Chem. 2020 Jan;12(1):48-55. doi: 10.1038/s41557-019-0366-y. Epub 2019 Nov 25.

Abstract

The field of synthetic biology has used the engineered assembly of synthetic gene networks to create a wide range of functions in biological systems. To date, gene-circuit-based sensors have primarily used optical proteins (for example, fluorescent, colorimetric) as reporter outputs, which has limited the potential to measure multiple distinct signals. Here we present an electrochemical interface that permits expanded multiplexed reporting for cell-free gene-circuit-based sensors. We have engineered a scalable system of reporter enzymes that cleave specific DNA sequences in solution, which results in an electrochemical signal when these newly liberated strands are captured at the surface of a nanostructured microelectrode. We describe the development of this interface and show its utility using a ligand-inducible gene circuit and toehold switch-based sensors by demonstrating the detection of multiple antibiotic resistance genes in parallel. This technology has the potential to expand the field of synthetic biology by providing an interface for materials, hardware and software.

Publication types

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

MeSH terms

  • Alkanesulfonates / chemistry
  • Azo Compounds / chemistry
  • DNA Restriction Enzymes / chemistry
  • DNA, Single-Stranded / chemistry*
  • DNA, Single-Stranded / genetics
  • DNA-Directed RNA Polymerases / chemistry
  • Drug Resistance, Multiple / genetics
  • Electrochemical Techniques / instrumentation
  • Electrochemical Techniques / methods*
  • Fluoresceins / chemistry
  • Gene Regulatory Networks*
  • Genes, MDR*
  • Methylene Blue / chemistry
  • Microelectrodes
  • Nucleic Acid Hybridization
  • Proof of Concept Study
  • RNA, Messenger / analysis
  • Viral Proteins / chemistry

Substances

  • Alkanesulfonates
  • Azo Compounds
  • BHQ-1 compound
  • DNA, Single-Stranded
  • Fluoresceins
  • RNA, Messenger
  • Viral Proteins
  • 6-carboxyfluorescein
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases
  • DNA Restriction Enzymes
  • Methylene Blue