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      A recombinase-based genetic circuit for heavy metal monitoring

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      Author(s)
      Akboğa, Doğuş
      Saltepe, Behide
      Date
      2022-02-16
      Source Title
      Biosensors
      Print ISSN
      20796374
      Publisher
      MDPI
      Volume
      12
      Issue
      2
      Pages
      1 - 10
      Language
      English
      Type
      Article
      Item Usage Stats
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      Abstract
      Rapid progress in the genetic circuit design enabled whole-cell biosensors (WCBs) to become prominent in detecting an extensive range of analytes with promise in many fields, from medical diagnostics to environmental toxicity assessment. However, several drawbacks, such as high background signal or low precision, limit WCBs to transfer from proof-of-concept studies to real-world applications, particularly for heavy metal toxicity monitoring. For an alternative WCB module design, we utilized Bxb1 recombinase that provides tight control as a switch to increase dose-response behavior concerning leakiness. The modularity of Bxb1 recombinase recognition elements allowed us to combine an engineered semi-specific heat shock response (HSR) promoter, sensitive to stress conditions including toxic ions such as cadmium, with cadmium resistance regulatory elements; a cadmium-responsive transcription factor and its cognitive promoter. We optimized the conditions for the recombinase-based cadmium biosensor to obtain increased fold change and shorter response time. This system can be expanded for various heavy metals to make an all-in-one type of WCB, even using semi-specific parts of a sensing system. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.
      Keywords
      Bxb1 recombinase
      Heavy metal detection
      Synthetic biology
      Toxicity
      Whole-cell biosensor
      Permalink
      http://hdl.handle.net/11693/112020
      Published Version (Please cite this version)
      https://dx.doi.org/10.3390/bios12020122
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      • Institute of Materials Science and Nanotechnology (UNAM) 2258
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