Toehold-mediated strand displacement (TMSD) is an isothermal switching process that enables the sequence-programmable and reversible conversion of DNA or RNA strands between single- and double-stranded conformations or other secondary structures. TMSD processes have already found widespread application in DNA nanotechnology, where they are used to drive DNA-based molecular devices or for the realization of synthetic biochemical computing circuits. Recently, researchers have started to employ TMSD also for the control of RNA-based gene regulatory processes in vivo, in particular in the context of synthetic riboregulators and conditional guide RNAs for CRISPR/Cas. Here, we provide a review over recent developments in this emerging field and discuss the opportunities and challenges for such systems in in vivo applications.
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