title: FRET-based genetically-encoded sensors for quantitative monitoring of metabolites.
abstract: Neighboring cells in the same tissue can exist in different states of dynamic activities. After genomics, proteomics and metabolomics, fluxomics is now equally important for generating accurate quantitative information on the cellular and sub-cellular dynamics of ions and metabolite, which is critical for functional understanding of organisms. Various spectrometry techniques are used for monitoring ions and metabolites, although their temporal and spatial resolutions are limited. Discovery of the fluorescent proteins and their variants has revolutionized cell biology. Therefore, novel tools and methods targeting sub-cellular compartments need to be deployed in specific cells and targeted to sub-cellular compartments in order to quantify the target-molecule dynamics directly. We require tools that can measure cellular activities and protein dynamics with sub-cellular resolution. Biosensors based on fluorescence resonance energy transfer (FRET) are genetically encoded and hence can specifically target sub-cellular organelles by fusion to proteins or targetted sequences. Since last decade, FRET-based genetically encoded sensors for molecules involved in energy production, reactive oxygen species and secondary messengers have helped to unravel key aspects of cellular physiology. This review, describing the design and principles of sensors, presents a database of sensors for different analytes/processes, and illustrate examples of application in quantitative live cell imaging.
authors.affiliation: Department of Biosciences, Jamia Millia Islamia, New Delhi, 110025, India. mmohsin1@jmi.ac.in.
authors.forename: Mohd
authors.initials: M
authors.lastname: Mohsin
authors.affiliation: Department of Botany, Aligarh Muslim University, Aligarh, 202002, India.
authors.forename: Altaf
authors.initials: A
authors.lastname: Ahmad
authors.affiliation: Department of Botany, Hamdard University, New Delhi, 110062, India.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Iqbal
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D064688:Metabolic Flux Analysis / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics
chemical_list: D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins
keywords: FRET; Fluorescent protein; Flux; Imaging; Metabolite; Sensor