Electroporation and microinjection successfully deliver single-stranded and duplex DNA into live cells as detected by FRET measurements

Rosemary A. Bamford, Zheng-yun Zhao, Neil A. Hotchin, Iain B. Styles, Gerard B. Nash, James H. R. Tucker, Roy Bicknell

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)
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Abstract

Förster resonance energy transfer (FRET) technology relies on the close proximity of two compatible fluorophores for energy transfer. Tagged (Cy3 and Cy5) complementary DNA strands forming a stable duplex and a doubly-tagged single strand were shown to demonstrate FRET outside of a cellular environment. FRET was also observed after transfecting these DNA strands into fixed and live cells using methods such as microinjection and electroporation, but not when using lipid based transfection reagents, unless in the presence of the endosomal acidification inhibitor bafilomycin. Avoiding the endocytosis pathway is essential for efficient delivery of intact DNA probes into cells.
Original languageEnglish
Article numbere95097
JournalPLoS ONE
Volume9
Issue number4
DOIs
Publication statusPublished - 22 Apr 2014

Keywords

  • Confocal laser microscopy
  • Confocal microscopy
  • Electroporation
  • Fluorescence resonance energy transfer
  • Lasers
  • Microinjection
  • Oligonucleotides
  • Transfection

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