Quantification of DNA binding to cell-surfaces by flow cytometry

Sharon L. McCoy, Frances A. Hausman, Mark E. Deffebach, Antony Bakke, Louise S. Merkens, Robert M. Bennett, Steven H. Hefeneider

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

DNA binding to cell-surfaces has been documented in several studies. The interaction of DNA with cells has been shown to have therapeutic potential as a non-viral form of gene delivery and DNA vaccination. Recently, bacterial DNA binding and internalization has been demonstrated in some cells to trigger secretion of cytokines and cell activation. Previous studies to quantify DNA binding to cells have used radiolabeled DNA. Here we report a non-radioactive assay for quantification of cell-surface DNA binding based on the isoparametric analysis of flow cytometric data as described by Chatelier et al., Embo J., 5 (1986) 1181. This assay has the advantage over previously used procedures in not employing radioactive material and being able to discriminate viable from non-viable cells that bind DNA. With the importance of understanding the interaction of DNA with cells, this assay may have application for the identification and characterization of reagents designed to either enhance or inhibit DNA binding to cells. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)141-146
Number of pages6
JournalJournal of Immunological Methods
Volume241
Issue number1-2
DOIs
StatePublished - Jul 31 2000

Keywords

  • DNA binding
  • Flow cytometry
  • Isoparametric analysis

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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    McCoy, S. L., Hausman, F. A., Deffebach, M. E., Bakke, A., Merkens, L. S., Bennett, R. M., & Hefeneider, S. H. (2000). Quantification of DNA binding to cell-surfaces by flow cytometry. Journal of Immunological Methods, 241(1-2), 141-146. https://doi.org/10.1016/S0022-1759(00)00205-2