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Novel Expression Vectors Enabling Induction of Gene Expression by Small-Interfering RNAs and MicroRNAs

Small-interfering RNAs and microRNAs are small ∼21–22 nucleotide long RNAs capable of posttranscriptional suppression of gene expression. The synthetic siRNAs are especially designed to target pre-specified genes and are common molecular biology tools. The miRNAs are endogenous regulators of gene ex...

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Autores principales: Harel, Liraz, Gefen, Nir, Carmi, Ofira, Orbach, Pini, Einat, Paz, Abitbol, Guy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4267845/
https://www.ncbi.nlm.nih.gov/pubmed/25514749
http://dx.doi.org/10.1371/journal.pone.0115327
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author Harel, Liraz
Gefen, Nir
Carmi, Ofira
Orbach, Pini
Einat, Paz
Abitbol, Guy
author_facet Harel, Liraz
Gefen, Nir
Carmi, Ofira
Orbach, Pini
Einat, Paz
Abitbol, Guy
author_sort Harel, Liraz
collection PubMed
description Small-interfering RNAs and microRNAs are small ∼21–22 nucleotide long RNAs capable of posttranscriptional suppression of gene expression. The synthetic siRNAs are especially designed to target pre-specified genes and are common molecular biology tools. The miRNAs are endogenous regulators of gene expression found in a wide variety of eukaryotes. miRNAs are currently utilized for diagnostics applications. Therapeutically, various miRNA-antagonizing tools are being explored and miRNAs are also utilized for cell-specific inhibition of the expression of gene therapy vectors harboring target sites for specific miRNAs. Here we show, for the first time, that siRNAs and miRNAs can be harnessed to induce gene expression. We designed special expression vectors in which target sites for artificial siRNAs or endogenous miRNAs are located between the transgene and an Upstream Inhibitory Region (UIR). We hypothesized that cleavage of the mRNA by siRNAs or miRNAs will separate the transgene from the UIR and the resulting uncapped mRNA will be capable of being translated. A UIR composed of seven open reading frames was found to be the most efficient inhibitor of the translation of the downstream transgene. We show that under such a configuration both artificial siRNAs and endogenous miRNAs were capable of inducing transgene expression. We show that using the diphtheria toxin A-chain gene, in combination with target sites for highly expressed miRNAs, specific induction of cell-death can be achieved, setting the stage for application to cancer therapy.
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spelling pubmed-42678452014-12-26 Novel Expression Vectors Enabling Induction of Gene Expression by Small-Interfering RNAs and MicroRNAs Harel, Liraz Gefen, Nir Carmi, Ofira Orbach, Pini Einat, Paz Abitbol, Guy PLoS One Research Article Small-interfering RNAs and microRNAs are small ∼21–22 nucleotide long RNAs capable of posttranscriptional suppression of gene expression. The synthetic siRNAs are especially designed to target pre-specified genes and are common molecular biology tools. The miRNAs are endogenous regulators of gene expression found in a wide variety of eukaryotes. miRNAs are currently utilized for diagnostics applications. Therapeutically, various miRNA-antagonizing tools are being explored and miRNAs are also utilized for cell-specific inhibition of the expression of gene therapy vectors harboring target sites for specific miRNAs. Here we show, for the first time, that siRNAs and miRNAs can be harnessed to induce gene expression. We designed special expression vectors in which target sites for artificial siRNAs or endogenous miRNAs are located between the transgene and an Upstream Inhibitory Region (UIR). We hypothesized that cleavage of the mRNA by siRNAs or miRNAs will separate the transgene from the UIR and the resulting uncapped mRNA will be capable of being translated. A UIR composed of seven open reading frames was found to be the most efficient inhibitor of the translation of the downstream transgene. We show that under such a configuration both artificial siRNAs and endogenous miRNAs were capable of inducing transgene expression. We show that using the diphtheria toxin A-chain gene, in combination with target sites for highly expressed miRNAs, specific induction of cell-death can be achieved, setting the stage for application to cancer therapy. Public Library of Science 2014-12-16 /pmc/articles/PMC4267845/ /pubmed/25514749 http://dx.doi.org/10.1371/journal.pone.0115327 Text en © 2014 Harel et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Harel, Liraz
Gefen, Nir
Carmi, Ofira
Orbach, Pini
Einat, Paz
Abitbol, Guy
Novel Expression Vectors Enabling Induction of Gene Expression by Small-Interfering RNAs and MicroRNAs
title Novel Expression Vectors Enabling Induction of Gene Expression by Small-Interfering RNAs and MicroRNAs
title_full Novel Expression Vectors Enabling Induction of Gene Expression by Small-Interfering RNAs and MicroRNAs
title_fullStr Novel Expression Vectors Enabling Induction of Gene Expression by Small-Interfering RNAs and MicroRNAs
title_full_unstemmed Novel Expression Vectors Enabling Induction of Gene Expression by Small-Interfering RNAs and MicroRNAs
title_short Novel Expression Vectors Enabling Induction of Gene Expression by Small-Interfering RNAs and MicroRNAs
title_sort novel expression vectors enabling induction of gene expression by small-interfering rnas and micrornas
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4267845/
https://www.ncbi.nlm.nih.gov/pubmed/25514749
http://dx.doi.org/10.1371/journal.pone.0115327
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