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  3. Unlocked Nucleic Acids for miRNA detection using two dimensional nano-graphene oxide

Unlocked Nucleic Acids for miRNA detection using two dimensional nano-graphene oxide

  • Biosens Bioelectron. 2017 Mar 15;89(Pt 1):551-557. doi: 10.1016/j.bios.2016.02.058.
Neil M Robertson 1,  Amy E Toscano 1,  Vincent E LaMantia 1,  Mustafa Salih Hizir 1,  Muhit Rana 1,  Mustafa Balcioglu 1,  Jia Sheng 2,  Mehmet V Yigit 3
Affiliations

Affiliations

  • 1 Department of Chemistry, University at Albany, State University of New York, 1400 Washington Avenue, Albany, NY 12222, United States.
  • 2 Department of Chemistry, University at Albany, State University of New York, 1400 Washington Avenue, Albany, NY 12222, United States; The RNA Institute, University at Albany, State University of New York, 1400 Washington Avenue, Albany, NY 12222, United States.
  • 3 Department of Chemistry, University at Albany, State University of New York, 1400 Washington Avenue, Albany, NY 12222, United States; The RNA Institute, University at Albany, State University of New York, 1400 Washington Avenue, Albany, NY 12222, United States. Electronic address: myigit@albany.edu.
PMID: 26944029 DOI: 10.1016/j.bios.2016.02.058
Abstract

In this study we have used Unlocked Nucleic Acids (UNAs) to discriminate a breast Cancer oncomiR from two Other miRNAs in the same RNA family using two-dimensional graphene oxide nanoassemblies. Fluorescently labeled single stranded probe strands and graphene oxide nanoassemblies have been used to detect miR-10b and discriminate it from miR-10a, which differs by only a single nucleotide (12th base from the 5' end), and miR-10c, which differs by only two nucleotides (12th and 16th Bases from the 5' end). We have determined the discrimination efficacy and detection capacity of a DNA probe with two inserted UNA monomers (UNA2), and compared it to the DNA probe with two purposefully inserted mutations (DNAM2) and full complementary sequence (DNAfull). We have observed that UNA2 is 50 times more powerful than DNAfull in discriminating miR-10b from miR-10c while generating an equally high fluorescence signal. This fluorescence signal was then further enhanced with the use of the highly specific Endonuclease dsDNase for an enzymatic amplification step. The results demonstrate that the underutilized UNAs have enormous potential for miRNA detection and offer remarkable discrimination efficacy over single and double mismatches.

Keywords

Biosensor; DNA; Graphene; MicroRNA (miRNA); Nanoparticle; Unlocked nucleic acid.

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