Fluorescent DNA nanotags featuring covalently attached intercalating dyes: Synthesis, antibody conjugation, and intracellular imaging

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

doi:10.1021/bc100485f

Fluorescent DNA nanotags featuring covalently attached intercalating dyes: Synthesis, antibody conjugation, and intracellular imaging

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

Biochemistry, Chemistry & Physics

Research output: Contribution to journal › Article › peer-review

25 Scopus citations

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.

Original language	English
Pages (from-to)	1491-1502
Number of pages	12
Journal	Bioconjugate Chemistry
Volume	22
Issue number	8
DOIs	https://doi.org/10.1021/bc100485f
State	Published - Aug 17 2011

Scopus Subject Areas

Biotechnology
Bioengineering
Biomedical Engineering
Pharmacology
Pharmaceutical Science
Organic Chemistry

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Stadler, A. L., Delos Santos, J. O., Stensrud, E. S., Dembska, A., Silva, G. L., Liu, S., Shank, N. I., Kunttas-Tatli, E., Sobers, C. J., Gramlich, P. M. E., Carell, T., Peteanu, L. A., McCartney, B. M., & Armitage, B. A. (2011). Fluorescent DNA nanotags featuring covalently attached intercalating dyes: Synthesis, antibody conjugation, and intracellular imaging. Bioconjugate Chemistry, 22(8), 1491-1502. https://doi.org/10.1021/bc100485f

@article{9438840f4a4e46989ddb2218a43a07c2,

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.",

author = "Stadler, {Andrea L.} and {Delos Santos}, {Junriz O.} and Stensrud, {Elizabeth S.} and Anna Dembska and Silva, {Gloria L.} and Shengpeng Liu and Shank, {Nathaniel I.} and Ezgi Kunttas-Tatli and Sobers, {Courtney J.} and Gramlich, {Philipp M.E.} and Thomas Carell and Peteanu, {Linda A.} and McCartney, {Brooke M.} and Armitage, {Bruce A.}",

year = "2011",

month = aug,

day = "17",

doi = "10.1021/bc100485f",

language = "English",

volume = "22",

pages = "1491--1502",

journal = "Bioconjugate Chemistry",

issn = "1043-1802",

publisher = "American Chemical Society",

number = "8",

}

Stadler, AL, Delos Santos, JO, Stensrud, ES, Dembska, A, Silva, GL, Liu, S, Shank, NI, Kunttas-Tatli, E, Sobers, CJ, Gramlich, PME, Carell, T, Peteanu, LA, McCartney, BM & Armitage, BA 2011, 'Fluorescent DNA nanotags featuring covalently attached intercalating dyes: Synthesis, antibody conjugation, and intracellular imaging', Bioconjugate Chemistry, vol. 22, no. 8, pp. 1491-1502. https://doi.org/10.1021/bc100485f

TY - JOUR

T1 - Fluorescent DNA nanotags featuring covalently attached intercalating dyes

T2 - Synthesis, antibody conjugation, and intracellular imaging

AU - Stadler, Andrea L.

AU - Delos Santos, Junriz O.

AU - Stensrud, Elizabeth S.

AU - Dembska, Anna

AU - Silva, Gloria L.

AU - Liu, Shengpeng

AU - Shank, Nathaniel I.

AU - Kunttas-Tatli, Ezgi

AU - Sobers, Courtney J.

AU - Gramlich, Philipp M.E.

AU - Carell, Thomas

AU - Peteanu, Linda A.

AU - McCartney, Brooke M.

AU - Armitage, Bruce A.

PY - 2011/8/17

Y1 - 2011/8/17

N2 - 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.

AB - 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.

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DO - 10.1021/bc100485f

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SN - 1043-1802

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JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

IS - 8

ER -

Fluorescent DNA nanotags featuring covalently attached intercalating dyes: Synthesis, antibody conjugation, and intracellular imaging

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