Department of Biosciences, Comsats University Islamabad Campus, Islamabad 45550, Pakistan.
Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada.
Biomolecules. 2021 Aug 26;11(9):1276. doi: 10.3390/biom11091276.
Genetically encoded biosensors based on engineered fluorescent proteins (FPs) are essential tools for monitoring the dynamics of specific ions and molecules in biological systems. Arsenic ion in the +3 oxidation state (As) is highly toxic to cells due to its ability to bind to protein thiol groups, leading to inhibition of protein function, disruption of protein-protein interactions, and eventually to cell death. A genetically encoded biosensor for the detection of As could potentially facilitate the investigation of such toxicity both in vitro and in vivo. Here, we designed and developed two prototype genetically encoded arsenic biosensors (GEARs), based on a bacterial As responsive transcriptional factor AfArsR from . We constructed FRET-based GEAR biosensors by insertion of AfArsR between FP acceptor/donor FRET pairs. We further designed and engineered single FP-based GEAR biosensors by insertion of AfArsR into GFP. These constructs represent prototypes for a new family of biosensors based on the ArsR transcriptional factor scaffold. Further improvements of the GEAR biosensor family could lead to variants with suitable performance for detection of As in various biological and environmental systems.
基于工程化荧光蛋白（FPs）的基因编码生物传感器是监测生物系统中特定离子和分子动态的重要工具。三价砷离子（As）由于能够与蛋白质巯基结合，从而抑制蛋白质功能、破坏蛋白质-蛋白质相互作用，最终导致细胞死亡，对细胞具有高度毒性。用于检测砷的基因编码生物传感器可能有助于体外和体内研究这种毒性。在这里，我们设计并开发了两种基于细菌砷反应性转录因子 AfArsR 的原型基因编码砷生物传感器（GEARs）。我们通过在 FP 受体/供体 FRET 对之间插入 AfArsR 构建了基于 FRET 的 GEAR 生物传感器。我们进一步通过将 AfArsR 插入 GFP 来设计和构建了基于单个 FP 的 GEAR 生物传感器。这些构建体代表了基于 ArsR 转录因子支架的新型生物传感器家族的原型。GEAR 生物传感器家族的进一步改进可能会导致具有适合检测各种生物和环境系统中砷性能的变体。
