Artificial microRNAs as siRNA shuttles: improved safety as compared to shRNAs in vitro and in vivo

Ryan L Boudreau; Inês Martins; Beverly L Davidson

doi:10.1038/mt.2008.231

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Artificial microRNAs as siRNA shuttles: improved safety as compared to shRNAs in vitro and in vivo

Journal article

Open access

Peer reviewed

Artificial microRNAs as siRNA shuttles: improved safety as compared to shRNAs in vitro and in vivo

Ryan L Boudreau, Inês Martins and Beverly L Davidson

Molecular therapy, Vol.17(1), pp.169-175

01/2009

DOI: 10.1038/mt.2008.231

PMCID: PMC2834985

PMID: 19002161

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Abstract

RNA interference (RNAi) provides a promising therapeutic approach to human diseases. However, data from recent reports demonstrate that short-hairpin RNAs (shRNAs) may cause cellular toxicity, and this warrants further investigation of the safety of using RNAi vectors. Earlier, in comparing hairpin-based RNAi vectors, we noted that shRNAs are highly expressed and yield an abundance of unprocessed precursors, whereas artificial microRNAs (miRNAs) are expressed at lower levels and are processed efficiently. We hypothesized that unprocessed shRNAs arise from the saturation of endogenous RNAi machinery, which poses likely a burden to cells. In this study, we tested that hypothesis by assessing the relative effects of shRNAs and artificial miRNAs on the processing and function of miRNAs. In competition assays, shRNAs disrupted miRNA biogenesis and function, whereas artificial miRNAs avoided this interference even when dosed to silence as effectively as shRNAs. We next compared the safety of these vectors in mouse cerebella, and found that shRNAs cause Purkinje cell neurotoxicity. By contrast, artificial miRNA expression was well tolerated, resulting in effective target gene silencing in Purkinje cells. These findings, together with data from earlier work in mouse striata, suggest that miRNA-based platforms are better suited for therapeutic silencing in the mammalian brain.

Immunohistochemistry

Cell Line

Blotting, Northern

Animals

Cerebellum - metabolism

Humans

Cerebellum - cytology

RNA, Small Interfering - adverse effects

Mice

MicroRNAs - adverse effects

Purkinje Cells - drug effects

Details

Title: Subtitle: Artificial microRNAs as siRNA shuttles: improved safety as compared to shRNAs in vitro and in vivo
Creators: Ryan L Boudreau - Department of Internal Medicine, University of Iowa, Iowa City, Iowa 52240, USA
Inês Martins
Beverly L Davidson
Resource Type: Journal article
Publication Details: Molecular therapy, Vol.17(1), pp.169-175
DOI: 10.1038/mt.2008.231
PMID: 19002161
PMCID: PMC2834985
NLM abbreviation: Mol Ther
ISSN: 1525-0016
eISSN: 1525-0024
Publisher: United States
Grant note: R01 HD044093 / NICHD NIH HHS P01 NS050210 / NINDS NIH HHS T32 HL007121 / NHLBI NIH HHS DK-54759 / NIDDK NIH HHS NS-50210 / NINDS NIH HHS HD-44093 / NICHD NIH HHS P30 DK054759 / NIDDK NIH HHS
Language: English
Date published: 01/2009
Academic Unit: Iowa Neuroscience Institute; Cardiovascular Medicine; Fraternal Order of Eagles Diabetes Research Center; Internal Medicine
Record Identifier: 9984070321202771

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