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Mechanistic analysis of the enhanced RNAi activity by 6-mCEPh-purine at the 5′ end of the siRNA guide strand

A key approach for improving siRNA efficacy is chemical modifications. Through an in silico screening of modifications at the 5′-end nucleobase of the guide strand, an adenine-derived compound called 6-(3-(2-carboxyethyl)phenyl)-purine (6-mCEPh-purine) was identified to improve the RNAi activity in...

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Autores principales: Brechin, Vincent, Shinohara, Fumikazu, Saito, Jun-ichi, Seitz, Hervé, Tomari, Yukihide
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7812867/
https://www.ncbi.nlm.nih.gov/pubmed/33177187
http://dx.doi.org/10.1261/rna.073775.119
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author Brechin, Vincent
Shinohara, Fumikazu
Saito, Jun-ichi
Seitz, Hervé
Tomari, Yukihide
author_facet Brechin, Vincent
Shinohara, Fumikazu
Saito, Jun-ichi
Seitz, Hervé
Tomari, Yukihide
author_sort Brechin, Vincent
collection PubMed
description A key approach for improving siRNA efficacy is chemical modifications. Through an in silico screening of modifications at the 5′-end nucleobase of the guide strand, an adenine-derived compound called 6-(3-(2-carboxyethyl)phenyl)-purine (6-mCEPh-purine) was identified to improve the RNAi activity in cultured human cells and in vivo mouse models. Nevertheless, it remains unclear how this chemical modification enhances the siRNA potency. Here, we used a series of biochemical approaches to quantitatively evaluate the effect of the 6-mCEPh-purine modification at each step in the assembly of the RNAi effector complex called RISC. We found that the modification improves the formation of mature RISC at least in two different ways, by fixing the loading orientation of siRNA duplexes and increasing the stability of mature RISC after passenger strand ejection. Our data will provide a molecular platform for further development of chemically modified siRNA drugs.
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spelling pubmed-78128672022-02-01 Mechanistic analysis of the enhanced RNAi activity by 6-mCEPh-purine at the 5′ end of the siRNA guide strand Brechin, Vincent Shinohara, Fumikazu Saito, Jun-ichi Seitz, Hervé Tomari, Yukihide RNA Article A key approach for improving siRNA efficacy is chemical modifications. Through an in silico screening of modifications at the 5′-end nucleobase of the guide strand, an adenine-derived compound called 6-(3-(2-carboxyethyl)phenyl)-purine (6-mCEPh-purine) was identified to improve the RNAi activity in cultured human cells and in vivo mouse models. Nevertheless, it remains unclear how this chemical modification enhances the siRNA potency. Here, we used a series of biochemical approaches to quantitatively evaluate the effect of the 6-mCEPh-purine modification at each step in the assembly of the RNAi effector complex called RISC. We found that the modification improves the formation of mature RISC at least in two different ways, by fixing the loading orientation of siRNA duplexes and increasing the stability of mature RISC after passenger strand ejection. Our data will provide a molecular platform for further development of chemically modified siRNA drugs. Cold Spring Harbor Laboratory Press 2021-02 /pmc/articles/PMC7812867/ /pubmed/33177187 http://dx.doi.org/10.1261/rna.073775.119 Text en © 2021 Brechin et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Article
Brechin, Vincent
Shinohara, Fumikazu
Saito, Jun-ichi
Seitz, Hervé
Tomari, Yukihide
Mechanistic analysis of the enhanced RNAi activity by 6-mCEPh-purine at the 5′ end of the siRNA guide strand
title Mechanistic analysis of the enhanced RNAi activity by 6-mCEPh-purine at the 5′ end of the siRNA guide strand
title_full Mechanistic analysis of the enhanced RNAi activity by 6-mCEPh-purine at the 5′ end of the siRNA guide strand
title_fullStr Mechanistic analysis of the enhanced RNAi activity by 6-mCEPh-purine at the 5′ end of the siRNA guide strand
title_full_unstemmed Mechanistic analysis of the enhanced RNAi activity by 6-mCEPh-purine at the 5′ end of the siRNA guide strand
title_short Mechanistic analysis of the enhanced RNAi activity by 6-mCEPh-purine at the 5′ end of the siRNA guide strand
title_sort mechanistic analysis of the enhanced rnai activity by 6-mceph-purine at the 5′ end of the sirna guide strand
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7812867/
https://www.ncbi.nlm.nih.gov/pubmed/33177187
http://dx.doi.org/10.1261/rna.073775.119
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