Abstract
Since the 1970s, the emergence and expansion of novel methods for calcium ion (Ca2+) detection have found diverse applications in vitro and in vivo across a series of model animal systems. Matched with advances in fluorescence imaging techniques, the improvements in the functional range and stability of various calcium indicators have significantly enhanced more accurate study of intracellular Ca2+ dynamics and its effects on cell signaling, growth, differentiation, and regulation. Nonetheless, the current limitations broadly presented by organic calcium dyes, genetically encoded calcium indicators, and calcium-responsive nanoparticles suggest a potential path toward more rapid optimization by taking advantage of a synthetic biology approach. This engineering-oriented discipline applies principles of modularity and standardization to redesign and interrogate endogenous biological systems. This review will elucidate how novel synthetic biology technologies constructed for eukaryotic systems can offer a promising toolkit for interfacing with calcium signaling and overcoming barriers in order to accelerate the process of Ca2+ detection optimization.
Highlights
Calcium ions (Ca2+ ) are an ancient and ubiquitous second messenger with diverse functions in cells in a wide array of cell types, including muscle cells, neurons, glial cells, immune cells, and oocytes
By integrating genetically encoded calcium indicators (GECIs) into the genomes of transgenic organisms, they can be continuously synthesized by the host cell machinery and reduce the temporal limitations of dyes rapidly being cleared from the cytosol [20,101]
Macrophages and osteogenic-macrophages were treated with the nanoprobe, and NIR fluorescence signals coincided with the green fluorescence signals of the calcium indicator Fluo-3 in co-stained cells, suggesting that this nanoprobe has the potential to detect and visualize intracellular Ca2+ levels
Summary
Calcium ions (Ca2+ ) are an ancient and ubiquitous second messenger with diverse functions in cells in a wide array of cell types, including muscle cells, neurons, glial cells, immune cells, and oocytes. Calcium signaling plays a key regulatory role across numerous physiological processes, including cell growth, gene regulation, neuronal synaptic activity, immune system function, hormone secretion signaling, fertilization, and the biomechanics of contraction [1]. The recent expansion of a toolkit which can incorporate RNAibased synthetic regulators (transcription factors, RNA-binding proteins) and engineered cell-signaling components have expanded the capacity to classify mammalian cells and regulate cell fate or morphology (see reviews [14] and [9]) These tools have current and potential applications with respect to targeting intracellular calcium signaling pathways with precise spatial and temporal control while reducing off-target effects. We go on to highlight the advantages and current limitations to establish a broad overview of the extent to which current synthetic biology tools have already been utilized to enhance study of calcium signaling and the enormous potential for synthetic biology to enhance and expand Ca2+ detection in animal systems even further
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