Fluorescent DNA Nanotags Featuring Covalently Attached Intercalating Dyes: Synthesis, Antibody Conjugation, and Intracellular Imaging
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- 作者：Andrea L. Stadler ; Junriz O. Delos Santos ; Elizabeth S. Stensrud ; Anna Dembska ; Gloria L. Silva ; Shengpeng Liu ; Nathaniel I. Shank ; Ezgi Kunttas-Tatli ; Courtney J. Sobers ; Philipp M. E. Gramlich ; Thomas Carell ; Linda A. Peteanu ; Brooke M. McCartney ; Bruce A. Armitage
- 刊名：Bioconjugate Chemistry
- 出版年：2011
- 出版时间：August 17, 2011
- 年：2011
- 卷：22
- 期：8
- 页码：1491-1502
- 全文大小：1163K
- 年卷期：v.22,no.8(August 17, 2011)
- ISSN：1520-4812
文摘
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.