Premium
A Quencher‐Free Linear Probe from Serinol Nucleic Acid with a Fluorescent Uracil Analogue
Author(s) -
Murayama Keiji,
Asanuma Hiroyuki
Publication year - 2020
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201900498
Subject(s) - nucleic acid , uracil , nucleobase , chemistry , thymine , fluorescence , rna , duplex (building) , context (archaeology) , peptide nucleic acid , nucleic acid analogue , förster resonance energy transfer , dna , nucleic acid structure , nucleotide , combinatorial chemistry , stereochemistry , nucleic acid thermodynamics , biochemistry , biology , physics , quantum mechanics , paleontology , gene
With the goal of developing a quencher‐free probe composed of an artificial nucleic acid, the fluorescent nucleobase analogue 5‐(perylenylethynyl)uracil ( Pe U), which was incorporated into totally artificial serinol nucleic acid (SNA) as a substitute for thymine, has been synthesized. In the context of a 12‐mer duplex with RNA, these fluorophores reduce duplex stability slightly compared with that of an SNA without Pe U modification; thus suggesting that structural distortion is not induced by the modification. If two Pe Us were incorporated at separate positions in an SNA, the fluorescent emission at λ ≈490 nm was clearly enhanced upon hybridization with complementary RNA. A quencher‐free SNA linear probe containing three Pe Us, each separated by six nucleobases, has been designed. Detection of target RNA with high sensitivity and discrimination of a single‐base mismatch has also been demonstrated.