3/20/2023 0 Comments Hydroxyl photolinkerĭue to numerous biological processes that are controlled at the genetic level, light-activated oligonucleotides for manipulating DNA, RNA, or protein function hold considerable promise. Optogenetics approaches, by which light can trigger the activity of photoresponsive ion channels in individual neurons, powerfully combine genetic targeting of neurons with imaging optics, resulting in photochemical control of cells within intact, living organisms. Additionally, a caged α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonist was developed to monitor surface-exposed AMPA receptors in individual Xenopus frog oocytes and in single cells in rat hippocampal cultures. microinjected 1-(4,5-dimethoxy-2-nitrophenyl)ethyl (DMNPE)-caged ATP into the central nervous system of Drosophila fruit flies to study the role of dopaminergic neurons in the control of movement. These advantages have led to the study of biological systems using “caged” molecules that are activated with near-UV, visible, or near-IR light. Light can penetrate noninvasively into cells and living tissues, is focused with high spatial and temporal resolution, and is orthogonal to most biological processes (particularly in mammals). Our understanding of how light energy can be harvested, transduced, and utilized in biological systems is rapidly expanding. In insects and plants, a flavoprotein named cryptochrome regulates the 24-h circadian clock in a light-dependent fashion, whereas at mid-ocean depths, where sunlight scarcely penetrates, most organisms resort to generating their own bioluminescence to distract predators or lure prey. More recently, protein photo-oxidation has been implicated in cataractogenesis. Well-studied photo-biochemical reactions include photosynthesis, DNA damage and repair mechanisms, and visual processes in the retina. Photochemistry is essential to life on Earth, as most organisms depend on photochemical processes for their existence. TIVA-isolated mRNA can be amplified and then analyzed using next-generation sequencing (RNA-seq). Expanding into new caged oligonucleotide applications, our lab has developed Transcriptome In Vivo Analysis (TIVA) technology, which provides the first noninvasive, unbiased method for isolating mRNA from single neurons in brain tissues. To improve capabilities for in vivo studies, we harnessed the rich inorganic photochemistry of ruthenium bipyridyl complexes to synthesize Ru-caged morpholino antisense oligonucleotides that remain inactive in zebrafish embryos until uncaged with visible light. This technique has been employed by our laboratory and others to photoregulate gene expression in cells and living organisms, typically using near UV-activated organic chromophores. This review focuses on caged oligonucleotides that incorporate site-specifically one or two photocleavable linkers, whose photolysis yields oligonucleotides with dramatic structural and functional changes. In the past decade, several new approaches for caging the structure and function of DNA and RNA oligonucleotides have been developed. Light-activated (“caged”) compounds have been widely employed for studying biological processes with high spatial and temporal control.
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