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Highly Fluorescent Conjugated Pyrenes in Nucleic Acid Probes: (Phenylethynyl)pyrenecarbonyl‐Functionalized Locked Nucleic Acids
Author(s) -
Astakhova Irina V.,
Korshun Vladimir A.,
Wengel Jesper
Publication year - 2008
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.200801077
Subject(s) - bathochromic shift , nucleic acid , pyrene , chemistry , fluorescence , monomer , dna , locked nucleic acid , oligonucleotide , conjugated system , rna , excimer , photochemistry , combinatorial chemistry , organic chemistry , biochemistry , polymer , physics , quantum mechanics , gene
In recent years, fluorescently labeled oligonucleotides have become a widely used tool in diagnostics, DNA sequencing, and nanotechnology. The recently developed (phenylethynyl)pyrenes are attractive dyes for nucleic acid labeling, with the advantages of long‐wave emission relative to the parent pyrene, high fluorescence quantum yields, and the ability to form excimers. Herein, the synthesis of six (phenylethynyl)pyrene‐functionalized locked nucleic acid (LNA) monomers M 1 – M 6 and their incorporation into DNA oligomers is described. Multilabeled duplexes display higher thermal stabilities than singly modified analogues. An increase in the number of phenylethynyl substituents attached to the pyrene results in decreased binding affinity towards complementary DNA and RNA and remarkable bathochromic shifts of absorption/emission maxima relative to the parent pyrene fluorochrome. This bathochromic shift leads to the bright fluorescence colors of the probes, which differ drastically from the blue emission of unsubstituted pyrene. The formation of intra‐ and interstrand excimers was observed for duplexes that have monomers M 1 – M 6 in both complementary strands and in numerous single‐stranded probes. If more phenylethynyl groups are inserted, the detected excimer signals become more intense. In addition, (phenylethynyl)pyrenecarbonyl–LNA monomers M 4 , M 5 , and M 6 proved highly useful for the detection of single mismatches in DNA/RNA targets.