Detection of intra-brain cytoplasmic 1 (BC1) long noncoding RNA using graphene oxide-fluorescence beacon detector

被引:13
作者
Kim, Mee Young [1 ]
Hwang, Do Won [1 ,2 ]
Li, Fangyuan [3 ]
Choi, Yoori [1 ]
Byun, Jung Woo [1 ]
Kim, Dongho [4 ]
Kim, Jee-Eun [4 ]
Char, Kookheon [3 ]
Lee, Dong Soo [1 ,2 ]
机构
[1] Seoul Natl Univ, Coll Med, Dept Nucl Med, Seoul, South Korea
[2] Seoul Natl Univ, Grad Sch Convergence Sci & Technol, Dept Mol Med & Biopharmaceut Sci, Seoul, South Korea
[3] Seoul Natl Univ, Natl Creat Res Initiat Ctr Intelligent Hybrids, Sch Chem & Biol Engn, WCU Program Chem Convergence Energy & Environm, Seoul, South Korea
[4] Genolution Inc, Asan Inst Life Sci, Seoul, South Korea
基金
新加坡国家研究基金会;
关键词
CELL; LOCALIZATION; SEQUENCE; SYSTEM; PROBES;
D O I
10.1038/srep22552
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Detection of cellular expression of long noncoding RNAs (lncRNAs) was elusive due to the ambiguity of exposure of their reactive sequences associated with their secondary/tertiary structures and dynamic binding of proteins around lncRNAs. Herein, we developed graphene-based detection techniques exploiting the quenching capability of graphene oxide (GO) flakes for fluorescent dye (FAM)-labeled single-stranded siRNAs and consequent un-quenching by their detachment from GO by matching lncRNAs. A brain cytoplasmic 1 (BC1) lncRNA expression was significantly decreased by a siRNA, siBC1-1. GO quenched the FAM-labeled siBC1-1 peptide nucleic acid (PNA) probe, and this quenching was recovered by BC1. While FAM-siBC1-1-PNA-GO complex transfected spontaneously mouse or human neural stem cells, fluorescence was recovered only in mouse cells having high BC1 expression. Fluorescent dye-labeled single-stranded RNA-GO probe could detect the reactive exposed nucleic acid sequence of a cytoplasmic lncRNA expressing in the cytoplasm, which strategy can be used as a detection method of lncRNA expression.
引用
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页数:7
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