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Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation

MicroRNA (miR) sponges containing miR binding sequences constitute a potentially powerful molecular therapeutic strategy. Recently, naturally occurring circular RNAs (circRNAs) were shown to function as efficient miR sponges in cancer cells. We hypothesized that synthetic circRNA sponges could achie...

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Autores principales: Liu, Xi, Abraham, John M., Cheng, Yulan, Wang, Zhixiong, Wang, Zhe, Zhang, Guanjun, Ashktorab, Hassan, Smoot, Duane T., Cole, Robert N., Boronina, Tatiana N., DeVine, Lauren R., Talbot, C. Conover, Liu, Zhengwen, Meltzer, Stephen J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6197335/
https://www.ncbi.nlm.nih.gov/pubmed/30326427
http://dx.doi.org/10.1016/j.omtn.2018.09.010
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author Liu, Xi
Abraham, John M.
Cheng, Yulan
Wang, Zhixiong
Wang, Zhe
Zhang, Guanjun
Ashktorab, Hassan
Smoot, Duane T.
Cole, Robert N.
Boronina, Tatiana N.
DeVine, Lauren R.
Talbot, C. Conover
Liu, Zhengwen
Meltzer, Stephen J.
author_facet Liu, Xi
Abraham, John M.
Cheng, Yulan
Wang, Zhixiong
Wang, Zhe
Zhang, Guanjun
Ashktorab, Hassan
Smoot, Duane T.
Cole, Robert N.
Boronina, Tatiana N.
DeVine, Lauren R.
Talbot, C. Conover
Liu, Zhengwen
Meltzer, Stephen J.
author_sort Liu, Xi
collection PubMed
description MicroRNA (miR) sponges containing miR binding sequences constitute a potentially powerful molecular therapeutic strategy. Recently, naturally occurring circular RNAs (circRNAs) were shown to function as efficient miR sponges in cancer cells. We hypothesized that synthetic circRNA sponges could achieve therapeutic loss-of-function targeted against specific miRs. Linear RNA molecules containing miR-21 binding sites were transcribed in vitro; after dephosphorylation and phosphorylation, circularization was achieved using 5′-3′ end-ligation by T4 RNA ligase 1. circRNA stability was assessed using RNase R and fetal bovine serum. Competitive inhibition of miR-21 activity by a synthetic circRNA sponge was assessed using luciferase reporter, cell proliferation, and cell apoptosis assays in three gastric cancer cell lines. circRNA effects on downstream proteins were also delineated by Tandem Mass Tag (TMT) labeling (data available via ProteomeXchange identifier PRIDE: PXD008584), followed by western blotting. We conclude that artificial circRNA sponges resistant to nuclease digestion can be synthesized using simple enzymatic ligation steps. These sponges inhibit cancer cell proliferation and suppress the activity of miR-21 on downstream protein targets, including the cancer protein DAXX. In summary, synthetic circRNA sponges represent a simple, effective, convenient strategy for achieving targeted loss of miR function in vitro, with potential future therapeutic application in human patients.
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spelling pubmed-61973352018-10-24 Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation Liu, Xi Abraham, John M. Cheng, Yulan Wang, Zhixiong Wang, Zhe Zhang, Guanjun Ashktorab, Hassan Smoot, Duane T. Cole, Robert N. Boronina, Tatiana N. DeVine, Lauren R. Talbot, C. Conover Liu, Zhengwen Meltzer, Stephen J. Mol Ther Nucleic Acids Article MicroRNA (miR) sponges containing miR binding sequences constitute a potentially powerful molecular therapeutic strategy. Recently, naturally occurring circular RNAs (circRNAs) were shown to function as efficient miR sponges in cancer cells. We hypothesized that synthetic circRNA sponges could achieve therapeutic loss-of-function targeted against specific miRs. Linear RNA molecules containing miR-21 binding sites were transcribed in vitro; after dephosphorylation and phosphorylation, circularization was achieved using 5′-3′ end-ligation by T4 RNA ligase 1. circRNA stability was assessed using RNase R and fetal bovine serum. Competitive inhibition of miR-21 activity by a synthetic circRNA sponge was assessed using luciferase reporter, cell proliferation, and cell apoptosis assays in three gastric cancer cell lines. circRNA effects on downstream proteins were also delineated by Tandem Mass Tag (TMT) labeling (data available via ProteomeXchange identifier PRIDE: PXD008584), followed by western blotting. We conclude that artificial circRNA sponges resistant to nuclease digestion can be synthesized using simple enzymatic ligation steps. These sponges inhibit cancer cell proliferation and suppress the activity of miR-21 on downstream protein targets, including the cancer protein DAXX. In summary, synthetic circRNA sponges represent a simple, effective, convenient strategy for achieving targeted loss of miR function in vitro, with potential future therapeutic application in human patients. American Society of Gene & Cell Therapy 2018-09-22 /pmc/articles/PMC6197335/ /pubmed/30326427 http://dx.doi.org/10.1016/j.omtn.2018.09.010 Text en © 2018 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Liu, Xi
Abraham, John M.
Cheng, Yulan
Wang, Zhixiong
Wang, Zhe
Zhang, Guanjun
Ashktorab, Hassan
Smoot, Duane T.
Cole, Robert N.
Boronina, Tatiana N.
DeVine, Lauren R.
Talbot, C. Conover
Liu, Zhengwen
Meltzer, Stephen J.
Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation
title Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation
title_full Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation
title_fullStr Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation
title_full_unstemmed Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation
title_short Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation
title_sort synthetic circular rna functions as a mir-21 sponge to suppress gastric carcinoma cell proliferation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6197335/
https://www.ncbi.nlm.nih.gov/pubmed/30326427
http://dx.doi.org/10.1016/j.omtn.2018.09.010
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