dc.contributor.authorGuo, Wei Mei
dc.contributor.authorKong, Kiat Whye
dc.contributor.authorBrown, Christopher John
dc.contributor.authorQuah, Soo Tng
dc.contributor.authorYeo, Hui Ling
dc.contributor.authorHoon, Shawn
dc.contributor.authorSeow, Yiqi
dc.date.accessioned2015-04-13T04:36:38Z
dc.date.available2015-04-13T04:36:38Z
dc.date.copyright2014en_US
dc.date.issued2014
dc.identifier.citationGuo, W. M., Kong, K. W., Brown, C. J., Quah, S. T., Yeo, H. L., Hoon, S., et al. (2014). Identification and characterization of an eIF4e DNA aptamer that inhibits proliferation with high throughput sequencing. Molecular therapy-nucleic acids, 3.en_US
dc.identifier.issn2162-2531en_US
dc.identifier.urihttp://hdl.handle.net/10220/25386
dc.description.abstractDevelopment of DNA aptamer screens that are both simple and informative can increase the success rate of DNA aptamer selection and induce greater adoption. High eIF4e levels contribute to malignancies, thus eIF4e presents itself as a valuable target for DNA aptamer-based inhibition screen. Here, we demonstrate a method for the rapid selection of looped DNA aptamers against eIF4e by combining negative selection and purification in a single step, followed by characterization with high throughput sequencing. The resulting aptamers show functional binding to eIF4e and inhibit translation initiation in biochemical assays. When transfected into cells, eIF4e aptamers cause a dramatic loss of cell proliferation in tumor cells as seen with eIF4e knockdown with antisense oligonucleotides, shRNAs, and siRNAs, hinting at therapeutic possibilities. With the large data set provided by high throughput sequencing, we demonstrate that selection happens in waves and that sequencing data can be used to infer aptamer structure. Lastly, we show that ligation of looped aptamers can enhance their functional effects. These results demonstrate a rapid protocol to screen and optimize aptamers against macromolecules of interest.en_US
dc.description.sponsorshipASTAR (Agency for Sci., Tech. and Research, S’pore)
dc.format.extent10 p.en_US
dc.language.isoenen_US
dc.relation.ispartofseriesMolecular therapy-nucleic acidsen_US
dc.rights© 2014. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/.en_US
dc.subjectDRNTU::Science::Biological sciences::Molecular biology
dc.titleIdentification and characterization of an eIF4e DNA aptamer that inhibits proliferation with high throughput sequencingen_US
dc.typeJournal Article
dc.contributor.schoolSchool of Biological Sciencesen_US
dc.identifier.doihttp://dx.doi.org/10.1038/mtna.2014.70
dc.description.versionPublished versionen_US


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