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Individually addressable and dynamic DNA gates for multiplexed cell sorting.
Dahotre, Shreyas N; Chang, Yun Min; Wieland, Andreas; Stammen, Samantha R; Kwong, Gabriel A.
Afiliación
  • Dahotre SN; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA 30332.
  • Chang YM; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA 30332.
  • Wieland A; Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA 30322.
  • Stammen SR; Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA 30322.
  • Kwong GA; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA 30332.
Proc Natl Acad Sci U S A ; 115(17): 4357-4362, 2018 04 24.
Article en En | MEDLINE | ID: mdl-29632190
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Linfocitos T CD4-Positivos / Linfocitos T CD8-positivos / Proliferación Celular / Citometría de Flujo / Coriomeningitis Linfocítica / Virus de la Coriomeningitis Linfocítica Límite: Animals Idioma: En Revista: Proc Natl Acad Sci U S A Año: 2018 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Linfocitos T CD4-Positivos / Linfocitos T CD8-positivos / Proliferación Celular / Citometría de Flujo / Coriomeningitis Linfocítica / Virus de la Coriomeningitis Linfocítica Límite: Animals Idioma: En Revista: Proc Natl Acad Sci U S A Año: 2018 Tipo del documento: Article Pais de publicación: Estados Unidos