A Quencher-Free Linear Probe from Serinol Nucleic Acid with a Fluorescent Uracil Analogue |
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Authors: | Dr. Keiji Murayama Prof. Dr. Hiroyuki Asanuma |
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Affiliation: | Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603 Japan |
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Abstract: | With the goal of developing a quencher-free probe composed of an artificial nucleic acid, the fluorescent nucleobase analogue 5-(perylenylethynyl)uracil (PeU), 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 PeU modification; thus suggesting that structural distortion is not induced by the modification. If two PeUs 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 PeUs, 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. |
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Keywords: | artificial nucleobases artificial oligonucleotides fluorescent probes nucleic acids perylenes |
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