Inhibitory Impact of 3 '-Terminal 2 '-0-Methylated Small Silencing RNA on Target-Primed Polymerization and Unbiased Amplified Quantification of the RNA in Arabidopsis thaliana
Chen, F; Fan, CH; Zhao, YX; Zhao, YX (reprint author), Xi An Jiao Tong Univ, Educ Minist, Sch Life Sci & Technol, Key Lab Biomed Informat Engn, Xianning West Rd, Xian 710049, Shaanxi, Peoples R China.
2015
发表期刊ANALYTICAL CHEMISTRY
ISSN0003-2700
卷号87期号:17页码:8758—8764
文章类型期刊文献
摘要3'-terminal 2'-O-methylation has been found in several kinds of small silencing RNA, regarded as a protective mechanism against enzymatic 3' -> 5' degradation and 3'-end uridylation. The influence of this modification on enzymatic polymerization, however, remains unknown. Herein, a systematic investigation is performed to explore this issue. We found these methylated small RNAs exhibited a suppression behavior in target-primed polymerization, revealing biased result for the manipulation of these small RNAs by conventional polymerization-based methodology. The related potential mechanism is investigated and discussed, which is probably ascribed to the big size of modified group and its close location to 3'-OH. Furthermore, two novel solutions each utilizing base-stacking hybridization and three-way junction structure have been proposed to realize unbiased recognition of small RNAs. On the basis of phosphorothioate against nicking, a creative amplified strategy, phosphorothioate-protected polymerization/binicking amplification, has also been developed for the unbiased quantification of methylated small RNA in Arabidopsis thaliana, demonstrating its promising potential for real sample analysis. Collectively, our studies uncover the polymerization inhibition by 3'-terminal 2'-O-methylated small RNAs with mechanistic discussion, and propose novel unbiased solutions for amplified quantification of small RNAs in real sample.
关键词Isothermal Amplification Sensitive Detection Ultrasensitive Detection 3-way Junction Microrna Dna Methylation Nanoparticles Hybridization Biogenesis
语种英语
WOS记录号WOS:000360773100025
引用统计
被引频次:14[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.sinap.ac.cn/handle/331007/24963
专题中科院上海应用物理研究所2011-2018年
通讯作者Zhao, YX (reprint author), Xi An Jiao Tong Univ, Educ Minist, Sch Life Sci & Technol, Key Lab Biomed Informat Engn, Xianning West Rd, Xian 710049, Shaanxi, Peoples R China.
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GB/T 7714
Chen, F,Fan, CH,Zhao, YX,et al. Inhibitory Impact of 3 '-Terminal 2 '-0-Methylated Small Silencing RNA on Target-Primed Polymerization and Unbiased Amplified Quantification of the RNA in Arabidopsis thaliana[J]. ANALYTICAL CHEMISTRY,2015,87(17):8758—8764.
APA Chen, F,Fan, CH,Zhao, YX,&Zhao, YX .(2015).Inhibitory Impact of 3 '-Terminal 2 '-0-Methylated Small Silencing RNA on Target-Primed Polymerization and Unbiased Amplified Quantification of the RNA in Arabidopsis thaliana.ANALYTICAL CHEMISTRY,87(17),8758—8764.
MLA Chen, F,et al."Inhibitory Impact of 3 '-Terminal 2 '-0-Methylated Small Silencing RNA on Target-Primed Polymerization and Unbiased Amplified Quantification of the RNA in Arabidopsis thaliana".ANALYTICAL CHEMISTRY 87.17(2015):8758—8764.
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