Acridone-tagged DNA as a new probe for DNA detection by fluorescence resonance energy transfer and for mismatch DNA recognition.
Acridone is highly fluorescent and stable against photodegradation, oxidation, and heat. It is also a small molecule with no charge, making it a promising fluorescent agent for use in a DNA probe. Thus, we have prepared 5'-terminal acridone-labeled DNAs by post-modification, and have examined their photophysical properties and their use as donors for a fluorescence resonance energy transfer (FRET) system in combination with a 3'-terminal dabcyl-tagged DNA as an acceptor, which can detect the target DNA by emission-quenching caused by FRET. The FRET with an acridone and dabcyl pair has been found to complement that with fluorescence and dabcyl and other fluorescence-quencher pairs. Significant amounts of quenching of the acridone emissions by guanine in the DNA were observed when guanine was close to acridone, which can be applied as a quencher-free probe for the detection of special sequence of DNA. The DNA bearing acridone at the C5 position of inner thymidine could discriminate the opposite T-T base mismatch, although enhancement of discrimination ability is needed for the practical use of SNP typing.
Department of Applied Chemistry and Chemical Biology, Gunma University, Kiryu, Gunma 376-8515, Japan.
Yasuhisa
Y
Hagiwara
Tomoya
T
Hasegawa
Atushi
A
Shoji
Masayasu
M
Kuwahara
Hiroaki
H
Ozaki
Hiroaki
H
Sawai
D000166:Acridines; D054831:Acridones; D020137:Base Pair Mismatch; D004247:DNA / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods; D015342:DNA Probes / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes; D006147:Guanine
D000166:Acridines; D054831:Acridones; D015342:DNA Probes; D005456:Fluorescent Dyes; D006147:Guanine; C041300:acridone; D004247:DNA