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Author Notes:

To whom correspondence should be addressed. Email: gkwong@gatech.edu

Author contributions: S.N.D. and G.A.K. designed research; S.N.D., Y.M.C., A.W., and S.R.S. performed research; A.W. contributed new reagents/analytic tools; S.N.D., Y.M.C., and G.A.K. analyzed data; and S.N.D. and G.A.K. wrote the paper.

We thank R. Ahmed (Emory) for insightful discussions.

The authors declare no conflict of interest.

Research Funding:

This work was funded by NIH Director’s New Innovator Award DP2HD091793 and by National Center for Advancing Translational Sciences of the NIH Award UL1TR000454.

S.N.D. is supported by NSF Graduate Research Fellowships Program Grant DGE-1650044 and NSF Integrative Graduate Education and Research Traineeship Grant 0965945.

Y.M.C. is supported by National Institute of General Medical Sciences of the NIH Award T32GM8169-30. G.A.K. holds a Career Award at the Scientific Interface from the Burroughs Wellcome Fund.

This work was performed in part at the Georgia Tech Institute for Electronics and Nanotechnology, a member of the National Nanotechnology Coordinated Infrastructure, which is supported by NSF Grant ECCS-1542174.

Keywords:

  • DNA nanotechnology
  • cell sorting
  • lymphocytic choriomeningitis virus
  • protein conjugates
  • strand displacement
  • Animals
  • CD4-Positive T-Lymphocytes
  • CD8-Positive T-Lymphocytes
  • Cell Proliferation
  • Flow Cytometry
  • Lymphocytic Choriomeningitis
  • Lymphocytic choriomeningitis virus
  • Mice

Individually addressable and dynamic DNA gates for multiplexed cell sorting

Tools:

Journal Title:

Proceedings of the National Academy of Sciences

Volume:

Volume 115, Number 17

Publisher:

, Pages 4357-4362

Type of Work:

Article | Final Publisher PDF

Abstract:

The ability to analyze and isolate cells based on the expression of specific surface markers has increased our understanding of cell biology and produced numerous applications for biomedicine. However, established cell-sorting platforms rely on labels that are limited in number due to biophysical constraints, such as overlapping emission spectra of fluorophores in FACS. Here, we establish a framework built on a system of orthogonal and extensible DNA gates for multiplexed cell sorting. These DNA gates label target cell populations by antibodies to allow magnetic bead isolation en masse and then selectively unlock by strand displacement to sort cells. We show that DNA gated sorting (DGS) is triggered to completion within minutes on the surface of cells and achieves target cell purity, viability, and yield equivalent to that of commercial magnetic sorting kits. We demonstrate multiplexed sorting of three distinct immune cell populations (CD8+, CD4+, and CD19+) from mouse splenocytes to high purity and show that recovered CD8+T cells retain proliferative potential and target cell-killing activity. To broaden the utility of this platform, we implement a double positive sorting scheme using DNA gates on peptide-MHC tetramers to isolate antigen-specific CD8+T cells from mice infected with lymphocytic choriomeningitis virus (LCMV). DGS can potentially be expanded with fewer biophysical constraints to large families of DNA gates for applications that require analysis of complex cell populations, such as host immune responses to disease.

Copyright information:

© 2018 National Academy of Sciences. All Rights Reserved.

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