Engineered Nuclear Hormone Receptor-Biosensors for Environmental Monitoring and Early Drug Discovery

Gierach, Izabela; Wood, David

doi:10.5772/16374

Cited by 1 publication

(1 citation statement)

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“…In contrast to other in vitro techniques, a further advantage of our system is that there is no need for any purification or complex enzyme stabilization steps . Furthermore, the modular nature of the fusion protein opens the possibility of optimizing the system by rapidly incorporating new reporter enzymes, while also expanding the RAPID biosensor to include diverse nuclear hormone receptors for human and animal applications …”

Section: Resultsmentioning

confidence: 99%

Cell-Free Protein Synthesis Approach to Biosensing hTRβ-Specific Endocrine Disruptors

et al. 2017

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Here we introduce a Rapid Adaptable Portable In vitro Detection biosensor platform (RAPID) for detecting ligands that interact with nuclear hormone receptors (NHRs). The RAPID platform can be adapted for field use, allowing rapid evaluation of endocrine disrupting chemicals (EDCs) presence or absence in environmental samples, and can also be applied for drug screening. The biosensor is based on an engineered, allosterically activated fusion protein, which contains the ligand binding domain from a target NHR (human thyroid receptor β in this work). In vitro expression of this protein using cell-free protein synthesis (CFPS) technology in the presence of an EDC leads to activation of a reporter enzyme, reported through a straightforward colorimetric assay output. In this work, we demonstrate the potential of this biosensor platform to be used in a portable "just-add-sample" format for near real-time detection. We also demonstrate the robust nature of the cell-free protein synthesis component in the presence of a variety of environmental and human samples, including sewage, blood, and urine. The presented RAPID biosensor platform is significantly faster and less labor intensive than commonly available technologies, making it a promising tool for detecting environmental EDC contamination and screening potential NHR-targeted pharmaceuticals.

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Section: Resultsmentioning

confidence: 99%