Locking-to-unlocking system is an efficient strategy to design DNA/silver nanoclusters (AgNCs) probe for human miRNAs

Pratik Shah, Suk Won Choi, Ho Jin Kim, Seok Keun Cho, Yong Joo Bhang, Moon Young Ryu, Peter Waaben Thulstrup, Morten Jannik Bjerrum, Seong Wook Yang

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

MicroRNAs (miRNAs), small non-coding RNA molecules, are important biomarkers for research and medical purposes. Here, we describe the development of a fast and simple method using highly fluorescent oligonucleotide-silver nanocluster probes (DNA/AgNCs) to efficiently detect specific miRNAs. Due to the great sequence diversity of miRNAs in humans and other organisms, a uniform strategy for miRNA detection is attractive. The concept presented is an oligonucleotide-based locking-to-unlocking system that can be endowed with miRNA complementarity while maintaining the same secondary structure. The locking-to-unlocking system is based on fold-back anchored DNA templates that consist of a cytosine-rich loop for AgNCs stabilization, an miRNA recognition site and an overlap region for hairpin stabilization. When an miRNA is recognized, fluorescence in the visible region is specifically extinguished in a concentration-dependent manner. Here, the exact composition of the fold-back anchor for the locking-to-unlocking system has been systematically optimized, balancing propensity for loop-structure formation, encapsulation of emissive AgNCs and target sensitivity. It is demonstrated that the applied strategy successfully can detect a number of cancer related miRNAs in RNA extracts from human cancer cell lines.

Original languageEnglish
Article number1377
JournalNucleic acids research
Volume44
Issue number6
DOIs
Publication statusPublished - 2015 Dec 17

All Science Journal Classification (ASJC) codes

  • Genetics

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    Shah, P., Choi, S. W., Kim, H. J., Cho, S. K., Bhang, Y. J., Ryu, M. Y., Thulstrup, P. W., Bjerrum, M. J., & Yang, S. W. (2015). Locking-to-unlocking system is an efficient strategy to design DNA/silver nanoclusters (AgNCs) probe for human miRNAs. Nucleic acids research, 44(6), [1377]. https://doi.org/10.1093/nar/gkv1377