방사선의학 웹진

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Abstract

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. 

Author information

​Kim MY1, Hwang do W1,2, Li F3, Choi Y1, Byun JW1, Kim D4, Kim JE4, Char K3, Lee DS1,2.

1Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Korea.

2Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, Korea.

3The National Creative Research Initiative Center for Intelligent Hybrids, School of Chemical &Biological Engineering, The WCU Program of Chemical Convergence for Energy &Environment, Seoul National University, Seoul, Korea.

4Genolution Inc., Asan Institute for Life Sciences, Korea.