Publications
2021
Bouhedda F., Ryckelynck M.
Compartmentalization-based technologies for in vitro selection and evolution of ribozymes and light-up RNA aptamers Book Chapter
In: & B. Masquida S. Müller, W. Winkler (Ed.): Ribozymes, vol. 28, pp. 721-738, John Wiley & Sons, Ltd, 2021.
Abstract | Links | BibTeX | Tags: catalytic RNAs, Droplet microfluidics, fluorescence, in vitro evolution, In vitro selection, light-up RNA aptamers, ribozymes, RYCKELYNCK, screening, Unité ARN
@inbook{nokey,
title = {Compartmentalization-based technologies for in vitro selection and evolution of ribozymes and light-up RNA aptamers},
author = {F. Bouhedda and M. Ryckelynck},
editor = {W. Winkler & B. Masquida S. Müller},
url = {https://onlinelibrary.wiley.com/doi/abs/10.1002/9783527814527.ch28},
doi = {10.1002/9783527814527.ch28},
year = {2021},
date = {2021-01-01},
booktitle = {Ribozymes},
volume = {28},
pages = {721-738},
publisher = {John Wiley & Sons, Ltd},
abstract = {Summary Catalytic RNAs, also known as ribozymes, are naturally found in every living cell where they can occupy functions as important as peptide bond formation catalysis or intron splicing just as two examples. Besides, ribozymes are thought to be very ancient molecules that might have been the key actors of the so-called RNA world, but they also hold great promise for plenty of modern applications. These features have stimulated the development of in vitro evolution methodologies aiming at characterizing existing but also isolate new artificial ribozymes. Whereas bulk approaches in which all the RNA sequences of library are assayed in a single reaction mixture may be efficient to select fast, single-turn-over and/or self-modifying catalysts, this format is less adapted to the isolation of multiple turnover trans-acting molecules. Instead, a compartmentalization approach in which each variant is isolated and assayed into an individual compartment is better suited. In this chapter, we review the different strategies available to perform such compartmentalization and that range from hand-made water-in-oil emulsion to more advanced microfluidic-assisted ultrahigh-throughput screening. We finally extend the applications scope of these technologies to other RNAs (i.e., light-up RNA aptamers) for which a functional screening may also reveal more efficient than more conventional bulk in vitro selections.},
keywords = {catalytic RNAs, Droplet microfluidics, fluorescence, in vitro evolution, In vitro selection, light-up RNA aptamers, ribozymes, RYCKELYNCK, screening, Unité ARN},
pubstate = {published},
tppubtype = {inbook}
}
Summary Catalytic RNAs, also known as ribozymes, are naturally found in every living cell where they can occupy functions as important as peptide bond formation catalysis or intron splicing just as two examples. Besides, ribozymes are thought to be very ancient molecules that might have been the key actors of the so-called RNA world, but they also hold great promise for plenty of modern applications. These features have stimulated the development of in vitro evolution methodologies aiming at characterizing existing but also isolate new artificial ribozymes. Whereas bulk approaches in which all the RNA sequences of library are assayed in a single reaction mixture may be efficient to select fast, single-turn-over and/or self-modifying catalysts, this format is less adapted to the isolation of multiple turnover trans-acting molecules. Instead, a compartmentalization approach in which each variant is isolated and assayed into an individual compartment is better suited. In this chapter, we review the different strategies available to perform such compartmentalization and that range from hand-made water-in-oil emulsion to more advanced microfluidic-assisted ultrahigh-throughput screening. We finally extend the applications scope of these technologies to other RNAs (i.e., light-up RNA aptamers) for which a functional screening may also reveal more efficient than more conventional bulk in vitro selections.