Synthesis of azobenzene-tethered DNA for reversible photo-regulation of DNA functions: hybridization and transcription.
A phosphoramidite monomer bearing an azobenzene is synthesized from D-threoninol. Using this monomer, azobenzene moieties can be introduced into oligodeoxyribonucleotide (DNA) at any position on a conventional DNA synthesizer. With this azobenzene-tethered DNA, formation and dissociation of a DNA duplex can be reversibly photo-regulated by cis-trans isomerization of the azobenzene. When the azobenzene takes a trans-form, a stable duplex is formed. After isomerization of the trans-azobenzene to its cis-form by UV-light irradiation (300 nm < lambda < 400 nm), the duplex can be dissociated into two strands. The duplex is reformed on photo-induced cis-trans isomerization (lambda > 400 nm). The introduction of azobenzenes into the T7 promoter at specific positions also efficiently and reversibly photo-regulates transcription by T7-RNA polymerase. The reversible regulation can be repeated many times without causing damage to the DNA or the azobenzene moiety. These procedures take approximately 10 d to complete.
Asanuma
Hiroyuki
H
Department of Molecular Design and Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan. asanuma@mol.nagoya-u.ac.jp
Liang
Xingguo
X
Nishioka
Hidenori
H
Matsunaga
Daijiro
D
Liu
Mingzhe
M
Komiyama
Makoto
M
D001391:Azo Compounds / Q000737:chemistry*; D004247:DNA / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification; D015342:DNA Probes; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry* / Q000235:genetics; D005786:Gene Expression Regulation; D007536:Isomerism; D008027:Light; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D010777:Photochemistry / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics; D013050:Spectrometry, Fluorescence; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics
D001391:Azo Compounds; D015342:DNA Probes; D009838:Oligodeoxyribonucleotides; D014764:Viral Proteins; D004247:DNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; C009850:azobenzene