Electroporation and microinjection successfully deliver single-stranded and duplex DNA into live cells as detected by FRET measurements.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 24755680.
- Also identified by DOI 10.1371/journal.pone.0095097 and PMC identifier 3995676.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- DNA, Single-Stranded
- Electroporation
- Fluorescence Resonance Energy Transfer
- Microinjections