Live Cell MicroRNA Imaging Using Cascade Hybridization Reaction

Abstract

Recent advances in RNA research have posed new directives in biology and chemistry to uncover the complex roles of ribonucleic acids in cellular processes. Innovative techniques to visualize native RNAs, particularly, short, low-abundance RNAs in live cells, can dramatically impact current research on the roles of RNAs in biology. Herein, we report a novel method for real-time, microRNA imaging inside live cells based on programmable oligonucleotide probes, which self-assemble through the Cascade Hybridization Reaction (CHR).

A smart ZnO@polydopamine-nucleic acid nanosystem for ultrasensitive live cell mRNA imaging by the target-triggered intracellular self-assembly of active DNAzyme nanostructures

“…These nonenzymatic signal amplification mechanisms, such as the cascade hybridization reaction (CHR), 5 cascade reaction 6 and hybridization chain reaction (HCR), 7 can significantly enhance the fluorescence signal produced by the primary hybridization events of hairpin DNA probes and targets. Despite the fact that these reports have featured the ultrasensitive imaging analysis of mRNA expression levels of live cells by enzyme-free amplification strategies, there is still a lack of effective nonenzymatic dual signal amplification techniques involving the use of catalytic nucleic acids (DNAzymes or ribozymes) for ultrasensitive live cell mRNA imaging.…”

A Graphene-enhanced imaging of microRNA with enzyme-free signal amplification of catalyzed hairpin assembly in living cells

“…The hybridization chain reactions performed in stringent conditions, and it has an important drawback that destabilizing nonspecific binding. Owing to recent advances in molecular engineering and programming of DNA nanotechnology, nucleic acid hybridization-based nanocircuits have allowed the detection of nucleic acid targets with tens-or even hundredsfold catalytic amplification (Cheglakov et al, 2015;Wu et al, 2015). Recently, the catalyzed hairpin assembly (CHA) has been shown to be an idea choice for amplification and has been widely used for detection of DNA, proteins and miRNA in buffer solution (Huang et al, 2012;Liu et al, 2015;Zhu et al, 2014).…”

Mechanisms for the epigenetic inheritance of stress response in single cells

“…It will be exciting to apply these reporters to the tracking of the long-term inheritance of these epigenetic marks in live cells. For detection of low-abundance miRNAs in live cells, a reporter has been developed based on programmable molecular hairpins that can self-assemble to produce FRET signal 58 . Finally, the combination of single-cell RNA-seq technology with cell-lineage tracking methods 59 can be very powerful to examine stress-induced global transcriptomic changes that occur in single cells and to track how these changes are inherited across cell lineages.…”