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RNA Interference

RNA Interference

RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...

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Updated: Mar 24, 2026

Determination of In Vitro and Cellular Turn-on Kinetics for Fluorogenic RNA Aptamers

Determination of In Vitro and Cellular Turn-on Kinetics for Fluorogenic RNA Aptamers

Published on: August 9, 2022

iSpinach: a fluorogenic RNA aptamer optimized for in vitro applications.

Alexis Autour¹, Eric Westhof¹, Michael Ryckelynck²

¹Architecture et Réactivité de l'ARN, Université de Strasbourg, Institut de biologie moléculaire et cellulaire du CNRS, 15 rue René Descartes, 67084, Strasbourg, France.

Nucleic Acids Research

|March 3, 2016

Summary

This summary is machine-generated.

Researchers engineered brighter, more stable RNA aptamers called iSpinach. This new molecule surpasses existing aptamers and enables sensitive, high-throughput assays for measuring ribozyme activity.

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Last Updated: Mar 24, 2026

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Area of Science:

Molecular Biology
Biochemistry
Biotechnology

Background:

Light-up RNA aptamers like Spinach are valuable tools in molecular biology.
Existing aptamers can suffer from limitations such as salt sensitivity and lower thermal stability.
There is a need for improved aptamers with enhanced properties for various applications.

Purpose of the Study:

To isolate and engineer brighter, more stable RNA aptamers.
To develop a novel aptamer, iSpinach, with superior folding and fluorescence properties.
To demonstrate the utility of iSpinach in a high-throughput fluorogenic assay.

Main Methods:

Random mutagenesis was employed to generate aptamer variants.
Microfluidic-assisted In Vitro Compartmentalization was used for high-throughput screening.
The engineered aptamer iSpinach was characterized for its folding and fluorescence properties.

Main Results:

Isolation of brighter Spinach RNA aptamer mutants with reduced salt sensitivity and increased thermal stability.
Development of iSpinach, an aptamer surpassing current fluorogenic aptamers based on DFHBI.
Demonstration of iSpinach in a sensitive, high-throughput assay for co-transcriptional measurement of ribozyme catalytic constant (kcat).

Conclusions:

iSpinach represents a significant advancement in aptamer technology.
The enhanced properties of iSpinach enable novel applications in biochemical assays.
This work provides a powerful new tool for studying RNA function and enzyme kinetics.