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. 2023 Feb 6;11:1118702. doi: 10.3389/fbioe.2023.1118702

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Copyright © 2023 Tellechea-Luzardo, Stiebritz and Carbonell.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The full stack biosensor development toolbox. (A) Computational tools can be used to determine the allosteric pockets of interaction between ligands and aTF. (B) Using these pockets as reference, docking computations can be carried out to assess the affinity of the aTF towards a library of putative ligand compounds. (C) The allosteric site computation can be validated using directed mutagenesis to evaluate changes in affinity or specificity. (D) ChIP technology allows researchers to determine the TFBSs of a newly discovered aTF. (E) Similarly, EMSA can be used to individually validate single TFBS. (F) SELEX can be used to artificially obtain new TFBS to the aTF. (G) Cell-free assays are a quick prototyping technology to test biosensor circuits once assembled. (H) Whole-cell biosensor experiments allow the characterisation of the biosensor circuit in conditions closer to in vivo applications.