Dendrimer-Based Signal Amplification of Click-Labelled DNA in Situ

Chembiochem. 2017 Sep 5;18(17):1716-1720. doi: 10.1002/cbic.201700209. Epub 2017 Jul 24.

Abstract

The in vivo incorporation of alkyne-modified bases into the genome of cells is today the basis for the efficient detection of cell proliferation. Cells are grown in the presence of ethinyl-dU (EdU), fixed and permeabilised. The incorporated alkynes are then efficiently detected by using azide-containing fluorophores and the CuI -catalysed alkyne-azide click reaction. For a world in which constant improvement in the sensitivity of a given method is driving diagnostic advancement, we developed azide- and alkyne-modified dendrimers that allow the establishment of sandwich-type detection assays that show significantly improved signal intensities and signal-to-noise ratios far beyond that which is currently possible.

Keywords: cell proliferation; click chemistry; dendrimers; fluorescence microscopy; high-throughput screening.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkynes / chemistry
  • Azides / chemistry
  • Catalysis
  • Cell Proliferation
  • Click Chemistry
  • Copper / chemistry
  • Cycloaddition Reaction
  • DNA / chemistry
  • DNA / metabolism*
  • Dendrimers / chemistry*
  • Fluorescent Dyes / chemistry*
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence

Substances

  • Alkynes
  • Azides
  • Dendrimers
  • Fluorescent Dyes
  • Copper
  • DNA