Cargando…
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...
Autores principales: | , , , , |
---|---|
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 |
_version_ | 1783637738352803840 |
---|---|
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. |
format | Online Article Text |
id | pubmed-7812867 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT brechinvincent mechanisticanalysisoftheenhancedrnaiactivityby6mcephpurineatthe5endofthesirnaguidestrand AT shinoharafumikazu mechanisticanalysisoftheenhancedrnaiactivityby6mcephpurineatthe5endofthesirnaguidestrand AT saitojunichi mechanisticanalysisoftheenhancedrnaiactivityby6mcephpurineatthe5endofthesirnaguidestrand AT seitzherve mechanisticanalysisoftheenhancedrnaiactivityby6mcephpurineatthe5endofthesirnaguidestrand AT tomariyukihide mechanisticanalysisoftheenhancedrnaiactivityby6mcephpurineatthe5endofthesirnaguidestrand |