Short-hairpin RNAs synthesized by T7 phage polymerase do not induce interferon

Short-hairpin RNAs synthesized by T7 phage polymerase do not induce interferon

RNA interference (RNAi) mediated by small-interfering RNAs (siRNAs) is a highly effective gene-silencing mechanism with great potential for gene-therapeutic applications. siRNA agents also exert non-target-related biological effects and toxicities, including immune-system stimulation. Specifically,...

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Main Authors: Gondai, Takuma, Yamaguchi, Kazuya, Miyano-Kurosaki, Naoko, Habu, Yuichirou, Takaku, Hiroshi
Format: Online
Language:English
Published: Oxford University Press 2008
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2241887/
id pubmed-2241887
recordtype oai_dc
spelling pubmed-22418872008-02-21 Short-hairpin RNAs synthesized by T7 phage polymerase do not induce interferon Gondai, Takuma Yamaguchi, Kazuya Miyano-Kurosaki, Naoko Habu, Yuichirou Takaku, Hiroshi Methods Online RNA interference (RNAi) mediated by small-interfering RNAs (siRNAs) is a highly effective gene-silencing mechanism with great potential for gene-therapeutic applications. siRNA agents also exert non-target-related biological effects and toxicities, including immune-system stimulation. Specifically, siRNA synthesized from the T7 RNA polymerase system triggers a potent induction of type-I interferon (IFN) in a variety of cells. Single-stranded RNA also stimulates innate cytokine responses in mammals. We found that pppGn (n = 2,3) associated with the 5′-end of the short-hairpin RNA (shRNA) from the T7 RNA polymerase system did not induce detectable amounts of IFN. The residual amount of guanine associated with the 5′-end and hairpin structures of the transcript was proportional to the reduction of the IFN response. Here we describe a T7 pppGn (n = 2,3) shRNA synthesis that does not induce the IFN response, and maintains the full efficacy of siRNA. Oxford University Press 2008-02 2008-01-21 /pmc/articles/PMC2241887/ /pubmed/18208841 http://dx.doi.org/10.1093/nar/gkm1043 Text en © 2008 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
repository_type Open Access Journal
institution_category Foreign Institution
institution US National Center for Biotechnology Information
building NCBI PubMed
collection Online Access
language English
format Online
author Gondai, Takuma
Yamaguchi, Kazuya
Miyano-Kurosaki, Naoko
Habu, Yuichirou
Takaku, Hiroshi
spellingShingle Gondai, Takuma
Yamaguchi, Kazuya
Miyano-Kurosaki, Naoko
Habu, Yuichirou
Takaku, Hiroshi
Short-hairpin RNAs synthesized by T7 phage polymerase do not induce interferon
author_facet Gondai, Takuma
Yamaguchi, Kazuya
Miyano-Kurosaki, Naoko
Habu, Yuichirou
Takaku, Hiroshi
author_sort Gondai, Takuma
title Short-hairpin RNAs synthesized by T7 phage polymerase do not induce interferon
title_short Short-hairpin RNAs synthesized by T7 phage polymerase do not induce interferon
title_full Short-hairpin RNAs synthesized by T7 phage polymerase do not induce interferon
title_fullStr Short-hairpin RNAs synthesized by T7 phage polymerase do not induce interferon
title_full_unstemmed Short-hairpin RNAs synthesized by T7 phage polymerase do not induce interferon
title_sort short-hairpin rnas synthesized by t7 phage polymerase do not induce interferon
description RNA interference (RNAi) mediated by small-interfering RNAs (siRNAs) is a highly effective gene-silencing mechanism with great potential for gene-therapeutic applications. siRNA agents also exert non-target-related biological effects and toxicities, including immune-system stimulation. Specifically, siRNA synthesized from the T7 RNA polymerase system triggers a potent induction of type-I interferon (IFN) in a variety of cells. Single-stranded RNA also stimulates innate cytokine responses in mammals. We found that pppGn (n = 2,3) associated with the 5′-end of the short-hairpin RNA (shRNA) from the T7 RNA polymerase system did not induce detectable amounts of IFN. The residual amount of guanine associated with the 5′-end and hairpin structures of the transcript was proportional to the reduction of the IFN response. Here we describe a T7 pppGn (n = 2,3) shRNA synthesis that does not induce the IFN response, and maintains the full efficacy of siRNA.
publisher Oxford University Press
publishDate 2008
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2241887/
_version_ 1611438956491046912

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