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Individually addressable and dynamic DNA gates for multiplexed cell sorting.

Shreyas N Dahotre1, Yun Min Chang1,2, Andreas Wieland2

  • 1Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA 30332.

Proceedings of the National Academy of Sciences of the United States of America
|April 11, 2018
PubMed
Summary
This summary is machine-generated.

Researchers developed DNA gated sorting (DGS), a novel cell isolation method. This technique uses DNA gates for highly pure and viable cell sorting, overcoming limitations of traditional platforms for biomedical research.

Keywords:
DNA nanotechnologycell sortinglymphocytic choriomeningitis virusprotein conjugatesstrand displacement

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Immunology

Background:

  • Established cell sorting methods face limitations due to the restricted number of labels and biophysical constraints.
  • Fluorescence-activated cell sorting (FACS) is hindered by overlapping emission spectra of fluorophores.

Purpose of the Study:

  • To establish a novel framework for multiplexed cell sorting using orthogonal and extensible DNA gates.
  • To overcome the limitations of existing cell-sorting platforms by enabling a higher number of labels.

Main Methods:

  • Developed a system of DNA gates that label target cell populations via antibodies for magnetic bead isolation.
  • Utilized strand displacement mechanism to selectively unlock DNA gates for cell sorting.
  • Demonstrated DNA gated sorting (DGS) on cell surfaces within minutes.

Main Results:

  • Achieved target cell purity, viability, and yield equivalent to commercial magnetic sorting kits.
  • Successfully performed multiplexed sorting of CD8+, CD4+, and CD19+ immune cells from mouse splenocytes.
  • Isolated antigen-specific CD8+ T cells using DNA gates on peptide-MHC tetramers.

Conclusions:

  • DNA gated sorting (DGS) offers a versatile and extensible platform for cell analysis and isolation.
  • The DGS platform overcomes biophysical constraints, enabling analysis of complex cell populations.
  • This technology has broad applications in immunology and understanding host immune responses to disease.