Detection of CAG repeat DNA sequences by pyrene-functionalized pyrrole-imidazole polyamides.
Five N-methylpyrrole-N-methylimidazole (Py-Im) polyamides possessing a fluorescent pyrene were synthesized by Fmoc solid-phase synthesis using Py/Im monomers and pyrenylbutyl-pyrrole monomer compound 9. The steady state fluorescence of conjugates 1-5 was examined in the presence and absence of (CAG)(12)-containing oligodeoxynucleotides (ODNs) 1 and 2. Of the conjugates, conjugate 1 showed no background emission around 470 nm in the absence of ODNs, and a clear increase of emission at 475 nm was observed upon addition of ODNs 1 and 2. The emission of conjugate 1 at 475 nm increased linearly with the concentration of ODN and the number of CAG repeats. The results indicate that conjugate 1 efficiently forms a pyrene excimer upon binding in the minor groove of DNA.
Bando
Toshikazu
T
Department of Chemistry, Graduate School of Science, Kyoto University, Sakyo, Kyoto 606-8502, Japan.
Fujimoto
Jun
J
Minoshima
Masafumi
M
Shinohara
Ken-Ichi
K
Sasaki
Shunta
S
Kashiwazaki
Gengo
G
Mizumura
Masatoshi
M
Sugiyama
Hiroshi
H
D001483:Base Sequence; D004247:DNA / Q000032:analysis*; D005453:Fluorescence; D007093:Imidazoles / Q000737:chemistry*; D009757:Nylons / Q000138:chemical synthesis / Q000737:chemistry*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011721:Pyrenes / Q000737:chemistry*; D011758:Pyrroles / Q000737:chemistry*; D017422:Sequence Analysis, DNA / Q000379:methods; D013050:Spectrometry, Fluorescence / Q000379:methods; D018911:Trinucleotide Repeats