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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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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. |
format | Online Article Text |
id | pubmed-4267845 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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