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DNA-Origami-Based Fluorescence Brightness Standards for Convenient and Fast Protein Counting in Live Cells

Nathan D. Williams, Ane Landajuela, Ravi Kiran Kasula, Wenjiao Zhou, John T. Powell, Zhiqun Xi, Farren J. Isaacs, View ORCID ProfileJulien Berro, Derek Toomre, View ORCID ProfileErdem Karatekin, View ORCID ProfileChenxiang Lin
doi: https://doi.org/10.1101/2020.09.20.305359
Nathan D. Williams
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
2Nanobiology Institute, Yale University, West Haven CT 06516, USA
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Ane Landajuela
2Nanobiology Institute, Yale University, West Haven CT 06516, USA
3Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06520, USA
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Ravi Kiran Kasula
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
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Wenjiao Zhou
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
2Nanobiology Institute, Yale University, West Haven CT 06516, USA
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John T. Powell
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
2Nanobiology Institute, Yale University, West Haven CT 06516, USA
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Zhiqun Xi
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
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Farren J. Isaacs
4Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520, USA
5Department of Biomedical Engineering, Yale University, New Haven, CT 06520, USA
6Systems Biology Institute, Yale University, West Haven, CT 06516, USA
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Julien Berro
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
2Nanobiology Institute, Yale University, West Haven CT 06516, USA
7Department of Molecular Biophysics and Biochemistry, New Haven, CT 06520, USA
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Derek Toomre
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
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Erdem Karatekin
2Nanobiology Institute, Yale University, West Haven CT 06516, USA
3Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06520, USA
7Department of Molecular Biophysics and Biochemistry, New Haven, CT 06520, USA
8Université de Paris, SPPIN – Saints-Pères Paris Institute for the Neurosciences, Centre National de la Recherche Scientifique (CNRS), F-75006 Paris, France
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Chenxiang Lin
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
2Nanobiology Institute, Yale University, West Haven CT 06516, USA
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  • For correspondence: chenxiang.lin@yale.edu
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Abstract

Fluorescence microscopy has been one of the most discovery-rich methods in biology. In the digital age, the discipline is becoming increasingly quantitative. Virtually all biological laboratories have access to fluorescence microscopes, but abilities to quantify biomolecule copy numbers are limited by the complexity and sophistication associated with current quantification methods. Here, we present DNA-origami-based fluorescence brightness standards for counting 5–300 copies of proteins in mammalian and bacterial cells, tagged with fluorescent proteins or organic dyes. Compared to conventional quantification techniques, our brightness standards are robust, straightforward to use, and compatible with nearly all fluorescence imaging applications, thereby providing a practical and versatile tool to quantify biomolecules via fluorescence microscopy.

Competing Interest Statement

The authors have declared no competing interest.

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Posted September 21, 2020.
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DNA-Origami-Based Fluorescence Brightness Standards for Convenient and Fast Protein Counting in Live Cells
Nathan D. Williams, Ane Landajuela, Ravi Kiran Kasula, Wenjiao Zhou, John T. Powell, Zhiqun Xi, Farren J. Isaacs, Julien Berro, Derek Toomre, Erdem Karatekin, Chenxiang Lin
bioRxiv 2020.09.20.305359; doi: https://doi.org/10.1101/2020.09.20.305359
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DNA-Origami-Based Fluorescence Brightness Standards for Convenient and Fast Protein Counting in Live Cells
Nathan D. Williams, Ane Landajuela, Ravi Kiran Kasula, Wenjiao Zhou, John T. Powell, Zhiqun Xi, Farren J. Isaacs, Julien Berro, Derek Toomre, Erdem Karatekin, Chenxiang Lin
bioRxiv 2020.09.20.305359; doi: https://doi.org/10.1101/2020.09.20.305359

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