Genetically encoded fluorescent indicators (also called sensors, biosensors, or reporters) are powerful tools for monitoring the dynamics of cellular physiology. They can be targeted to specific cellular or subcellular regions by gene promoters, expression systems, protein recombination schemes or localization sequences, etc. A genetically encoded fluorescent indicator contains a sensing element that selectively binds the target molecule and a reporter unit that converts the interaction into a detectable signal. Most often, indicators produce chimeric proteins for visualizing the biochemical activity processes of the cells. The DNA sequence of the chimeric protein is usually located downstream of the gene promoter, which can be activated by the target analyte. Fluorescent proteins are the most used detectors, commonly red fluorescent protein (RFP) or green fluorescent protein (GFP), because of the ease of signal registration and the wide dynamic range that allows recording of small fluctuations in response to the target signal. In addition, these fluorescent proteins lack cytotoxicity and therefore do not affect the normal physiological processes of cells, while real-time monitoring can be performed.
Due to their structural features, genetically encoded fluorescent indicators have a wide range of detection capabilities. Genetically encoded fluorescent indicators are a reliable tool for pollutant analysis, drug screening, and investigation of toxicants. The advantage of genetically encoded fluorescent indicators over other detection methods is that, in addition to qualitative detection of target molecules, they provide a rapid and sensitive detection method for monitoring under natural physiological or pathological conditions. Typically, genetically encoded fluorescent indicators are cultivated in a population of cells, so the data can be applied to understand cell interactions. Genetically encoded fluorescent indicators can respond to fluctuations in the normal physiological activities of cells and tissues caused by external and internal stimuli. Genetically encoded fluorescent indicators combined with gene editing can be used for the visualization of signaling pathways and their interference, biomolecular and organelle interactions, and cell culture monitoring. Genetically encoded indicators with sensitivity, specificity, and compatibility have been explored and developed to study various biological process occurring in cells. Neurons, myocytes, and pancreatic cells are particularly widely used in biomedical research. In these cells, it is possible to measure the electrical response of cells when exposed to a stimulus by genetically encoded fluorescent voltage indicators. Genetically encoded indicators are widely used to record neural activity, including neurotransmitter release, calcium ion dynamics, and membrane potential fluctuations.
Genetically encoded fluorescent indicators are cheaper, easier to use, more specific, faster, and able to work in real time than traditional methods for determining biomolecules in living systems. Amerigo Scientific offers easy-to-operate genetically encoded fluorescent indicators with stable delivery and expression systems for studying biological processes in live cells.
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