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Nanomechanics and co-transcriptional folding of Spinach and Mango
Nanomechanics and co-transcriptional folding of Spinach and Mango
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Nanomechanics and co-transcriptional folding of Spinach and Mango
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Nanomechanics and co-transcriptional folding of Spinach and Mango
Nanomechanics and co-transcriptional folding of Spinach and Mango
Journal Article

Nanomechanics and co-transcriptional folding of Spinach and Mango

2019
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Overview
Recent advances in fluorogen-binding “light-up” RNA aptamers have enabled protein-free detection of RNA in cells. Detailed biophysical characterization of folding of G-Quadruplex (GQ)-based light-up aptamers such as Spinach, Mango and Corn is still lacking despite the potential implications on their folding and function. In this work we employ single-molecule fluorescence-force spectroscopy to examine mechanical responses of Spinach2, i MangoIII and MangoIV. Spinach2 unfolds in four discrete steps as force is increased to 7 pN and refolds in reciprocal steps upon force relaxation. In contrast, GQ-core unfolding in i MangoIII and MangoIV occurs in one discrete step at forces >10 pN and refolding occurred at lower forces showing hysteresis. Co-transcriptional folding using superhelicases shows reduced misfolding propensity and allowed a folding pathway different from refolding. Under physiologically relevant pico-Newton levels of force, these aptamers may unfold in vivo and subsequently misfold. Understanding of the dynamics of RNA aptamers will aid engineering of improved fluorogenic modules for cellular applications. Light-up aptamers are widely used for fluorescence visualization of non-coding RNA in vivo. Here the authors employ single-molecule fluorescence-force spectroscopy to characterize the mechanical responses of the G-Quadruplex based light-up aptamers Spinach2, i MangoIII and MangoIV, which is of interest for the development of improved fluorogenic modules for imaging applications.