title: Fluorescent DNA nanotags featuring covalently attached intercalating dyes: synthesis, antibody conjugation, and intracellular imaging.
abstract: We have synthesized fluorescent DNA duplexes featuring multiple thiazole orange (TO) intercalating dyes covalently attached to the DNA via a triazole linkage. The intercalating dyes stabilize the duplex against thermal denaturation and show bright fluorescence in the green region of the spectrum. The emission color can be changed to orange or red by addition of energy-accepting Cy3 or Cy5 dyes attached covalently to the DNA duplex. The dye-modified DNA duplexes were then attached to a secondary antibody for intracellular fluorescence imaging of centrosomes in Drosophila embryos. Bright fluorescent foci were observed at the centrosomes in both the donor (TO) and acceptor (Cy5) channels, because the energy transfer efficiency is moderate. Monitoring the Cy5 emission channel significantly minimized the background signal because of the large shift in emission wavelength allowed by energy transfer.
authors.affiliation: Department of Chemistry, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Andrea L
authors.initials: AL
authors.lastname: Stadler
authors.forename: Junriz O
authors.initials: JO
authors.lastname: Delos Santos
authors.forename: Elizabeth S
authors.initials: ES
authors.lastname: Stensrud
authors.forename: Anna
authors.initials: A
authors.lastname: Dembska
authors.forename: Gloria L
authors.initials: GL
authors.lastname: Silva
authors.forename: Shengpeng
authors.initials: S
authors.lastname: Liu
authors.forename: Nathaniel I
authors.initials: NI
authors.lastname: Shank
authors.forename: Ezgi
authors.initials: E
authors.lastname: Kunttas-Tatli
authors.forename: Courtney J
authors.initials: CJ
authors.lastname: Sobers
authors.forename: Philipp M E
authors.initials: PM
authors.lastname: Gramlich
authors.forename: Thomas
authors.initials: T
authors.lastname: Carell
authors.forename: Linda A
authors.initials: LA
authors.lastname: Peteanu
authors.forename: Brooke M
authors.initials: BM
authors.lastname: McCartney
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Armitage
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D002232:Carbocyanines; D018385:Centrosome / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D003952:Diagnostic Imaging / Q000379:methods*; D004330:Drosophila / Q000166:cytology; D004735:Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D018796:Immunoconjugates / Q000737:chemistry*; D007364:Intercalating Agents; D011804:Quinolines
chemical_list: D052160:Benzothiazoles; D002232:Carbocyanines; D005456:Fluorescent Dyes; D018796:Immunoconjugates; D007364:Intercalating Agents; D011804:Quinolines; C085321:cyanine dye 5; C051000:thiazole orange; D004247:DNA