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Programming in Situ Immunofluorescence Intensities through Interchangeable Reactions of Dynamic DNA Complexes

Authors

  • Jan Zimak,

    1. Departments of Bioengineering and Chemistry, Rice University, BRC 520, 6500 Main St, Houston, Texas, 77030 (USA)
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  • Dr. Ryan M. Schweller,

    1. Departments of Bioengineering and Chemistry, Rice University, BRC 520, 6500 Main St, Houston, Texas, 77030 (USA)
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  • Dr. Dzifa Y. Duose,

    1. Departments of Bioengineering and Chemistry, Rice University, BRC 520, 6500 Main St, Houston, Texas, 77030 (USA)
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  • Dr. Walter N. Hittelman,

    1. Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, 1901 East Road, Unit 1950, Houston, Texas, 77054 (USA)
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  • Dr. Michael R. Diehl

    Corresponding author
    1. Departments of Bioengineering and Chemistry, Rice University, BRC 520, 6500 Main St, Houston, Texas, 77030 (USA)
    • Departments of Bioengineering and Chemistry, Rice University, BRC 520, 6500 Main St, Houston, Texas, 77030 (USA)
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Abstract

The regulation of antibody reporting intensities is critical to various in situ fluorescence-imaging analyses. Although such control is often necessary to visualize sparse molecular targets, the ability to tune marker intensities is also essential for highly multiplexed imaging strategies in which marker reporting levels must be tuned both to optimize dynamic detection ranges and to minimize crosstalk between different signals. Existing chemical amplification approaches generally lack such control. Here, we demonstrate that linear and branched DNA complexes can be designed to function as interchangeable building blocks that can be assembled into organized, fluorescence-reporting complexes. We show that the ability to program DNA-strand-displacement reactions between these complexes offers new opportunities to deterministically tune the number of dyes that are coupled to individual antibodies in order both to increase and controllably balance marker reporting levels within fixed cells.

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