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The ability of DNA and RNA to fold into precise nanostructures has been used in a variety of biological and biomedical fields.
the precise self-assembled nanostructures of these nucleic acids create a good platform for the development of programmable molecular identification tools.
over the past decade, a complex set of nucleic acid-based circuits has been designed and applied to bioanalysis and detection.
one of the catalytic hairpin components (CHA) has proven to be one of them.
these enzyme-free and other temperature CHA circuits have the advantages of fast and effective signal amplification, low background and high turnover.
DNA circuits based on these highly sensitive and targeted specific CHA reactions have been applied to a variety of in vitro cell-free analyses, including nucleic acids, small molecules, and protein detection and quantitative analysis.
but most DNA-based circuits have the disadvantage of biological delivery and degradation difficulties.
on the other hand, RNA molecules can be genetically encoded and transcribed within the life system.
therefore, RNA-based circuits and devices should have great potential for intracellular applications.
: Professor Yu Mingxu of the University of Massachusetts and others recently reported on the genetic code of an RNA-based catalytic hairpin assembly circuit for sensitive RNA imaging in living cells.
the split form of broccoli, a fluorescent RNA organism, is used as a reporting molecule.
a target RNA can catalyze the fluorescence of dozens to hundreds of Broccoli, sensitively targeting RNA detection.
and further optimize the circuits they design, which can be easily programmed to target various RNA sequences for imaging.
design principle opens up a new stage for the development of various gene-coded RNA circuits for cell applications.
research published in the prestigious international journal JACS with the title "Genetically Encoded Catalytic Hairpin Assembly for Sensitive RNA Imaging in Live Cells".
: Figure I, CHARGE's circuit schematic II, different sequence design optimization CHARGE's signal-to-noise ratio figure III, D2 CHARGE system's body appearance signs (a) with or without target, The emission spectrum of the Scoreli and CHARGE circuits;
, live BL21 (DE3) cells for confocal fluorescence imaging, modular CHARGE system can detect a variety of RNA targets (a) based on molecular beacon smnob-based target detection schematics;
Figure VI, HHR-based soobases regulate cell RNA levels (a) the mesobases-induced HHR self-cutting and target RNA release schematics; (b) in vitro fluorescence assays of different concentrations of theteabase; (c) add different concentrations of the tea base 2 h, 10-base-long-long-long-to-long inhibitors incorporated in the HHR system 10% denatured PAGE gel signs; The
Summary study introduced a new gene-coded RNA circuit that can detect cell RNA targets with high sensitivity.
these CHARGE circuits can be easily programmed to image various RNA targets in living cells.
even in bacterial cells, current circuits have been confirmed, and CHARGE does have the potential for eukaryotic cell research.
a common application platform similar to the catalytic hairclip assembly circuit for in vitro analysis, a variety of CHARGEcans can be developed to further detect other cell RNAs, proteins and small molecules, as well as switches for genetic regulation.
the study opens up new ways to develop cell applications using genetically coded RNA circuits.
documentary link: Genetically Encoded Catalytic Hairpin Assembly for Sensitive RNA Imaging in Live Cells (JACS, 2018, DOI: 10.1021/jacs.8b03956) Source: Little Fat Paper Material Man.