@article{12de320e1134481ca5d4a2a4ff69f0ca,
title = "Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation",
abstract = "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.",
keywords = "gastric carcinoma, microRNA loss-of-function, microRNA sponge, molecular therapy, synthetic circular RNA",
author = "Xi Liu and Abraham, {John M.} and Yulan Cheng and Zhixiong Wang and Zhe Wang and Guanjun Zhang and Hassan Ashktorab and Smoot, {Duane T.} and Cole, {Robert N.} and Boronina, {Tatiana N.} and DeVine, {Lauren R.} and Talbot, {C. Conover} and Zhengwen Liu and Meltzer, {Stephen J.}",
note = "Funding Information: S.J.M. was supported by NIH grants CA190040, CA211457, and DK118250 and the Emerson Research Foundation; X.L. is supported by the Key Science and Technology Program of Shaanxi Province, China grant 2015SF128; G.Z. is supported by the International Scientific and Technological Cooperation and Exchange Program of Shaanxi Province, China grant 2015KW-030; and R.N.C. and T.N.B. were supported by Hopkins Digestive Diseases Basic and Translational Research Core Center (NIDDK center) grant P30 DK089502. S.J.M. is the Harry and Betty Myerberg Professor of Gastroenterology and American Cancer Society Clinical Research Professor. Funding Information: S.J.M. was supported by NIH grants CA190040 , CA211457 , and DK118250 and the Emerson Research Foundation ; X.L. is supported by the Key Science and Technology Program of Shaanxi Province, China grant 2015SF128 ; G.Z. is supported by the International Scientific and Technological Cooperation and Exchange Program of Shaanxi Province, China grant 2015KW-030 ; and R.N.C. and T.N.B. were supported by Hopkins Digestive Diseases Basic and Translational Research Core Center (NIDDK center) grant P30 DK089502 . S.J.M. is the Harry and Betty Myerberg Professor of Gastroenterology and American Cancer Society Clinical Research Professor. Publisher Copyright: {\textcopyright} 2018 The Authors",
year = "2018",
month = dec,
day = "7",
doi = "10.1016/j.omtn.2018.09.010",
language = "English (US)",
volume = "13",
pages = "312--321",
journal = "Molecular Therapy - Nucleic Acids",
issn = "2162-2531",
publisher = "Nature Publishing Group",
}