Design and Optimization of a Cell-Free Atrazine Biosensor

Adam D. Silverman, Umut Akova, Khalid K. Alam, Michael C. Jewett*, Julius B. Lucks

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

91 Scopus citations

Abstract

Recent advances in cell-free synthetic biology have spurred the development of in vitro molecular diagnostics that serve as effective alternatives to whole-cell biosensors. However, cell-free sensors for detecting manmade organic water contaminants such as pesticides are sparse, partially because few characterized natural biological sensors can directly detect such pollutants. Here, we present a platform for the cell-free detection of one critical water contaminant, atrazine, by combining a previously characterized cyanuric acid biosensor with a reconstituted atrazine-to-cyanuric acid metabolic pathway composed of several protein-enriched bacterial extracts mixed in a one pot reaction. Our cell-free sensor detects atrazine within an hour of incubation at an activation ratio superior to previously reported whole-cell atrazine sensors. We also show that the response characteristics of the atrazine sensor can be tuned by manipulating the ratios of enriched extracts in the cell-free reaction mixture. Our approach of utilizing multiple metabolic steps, encoded in protein-enriched cell-free extracts, to convert a target of interest into a molecule that can be sensed by a transcription factor is modular. Our work thus serves as an effective proof-of-concept for a scheme of "metabolic biosensing", which should enable rapid, field-deployable detection of complex organic water contaminants.

Original languageEnglish (US)
Pages (from-to)671-677
Number of pages7
JournalACS synthetic biology
Volume9
Issue number3
DOIs
StatePublished - Mar 20 2020

Funding

This work was supported by the Air Force Research Laboratory Center of Excellence for Advanced Bioprogrammable Nanomaterials (C-ABN) Grant FA8650-15-2-5518 (to M.C.J. and J.B.L.), the U.S. Defense Advanced Research Projects Agency’s (DARPA) Living Foundries program award HR0011-15-C-0084, the David and Lucile Packard Foundation (to M.C.J.), the Camille Dreyfus Teacher-Scholar Program (to M.C.J. and J.B.L.), an NSF CAREER Award (Grant 1452441 to J.B.L.), and Searle Funds at the Chicago Community Trust (to J.B.L.). A.D.S. was supported in part by the National Institutes of Health Training Grant (T32GM008449) through Northwestern University’s Biotechnology Training Program.

Keywords

  • TX-TL
  • atrazine
  • biosensor
  • cell-free
  • cyanuric acid
  • metabolism
  • synthetic biology
  • transcription factor

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Fingerprint

Dive into the research topics of 'Design and Optimization of a Cell-Free Atrazine Biosensor'. Together they form a unique fingerprint.

Cite this