title: Electroporation and microinjection successfully deliver single-stranded and duplex DNA into live cells as detected by FRET measurements.
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.
authors.affiliation: PSIBS Doctoral Training Centre, University of Birmingham, Birmingham, United Kingdom.
authors.forename: Rosemary A
authors.initials: RA
authors.lastname: Bamford
authors.affiliation: School of Chemistry, University of Birmingham, Birmingham, United Kingdom.
authors.forename: Zheng-yun
authors.initials: ZY
authors.lastname: Zhao
authors.affiliation: School of Biosciences, University of Birmingham, Birmingham, United Kingdom.
authors.forename: Neil A
authors.initials: NA
authors.lastname: Hotchin
authors.affiliation: School of Computer Science, University of Birmingham, Birmingham, United Kingdom.
authors.forename: Iain B
authors.initials: IB
authors.lastname: Styles
authors.affiliation: Centre for Cardiovascular Sciences, School of Clinical and Experimental Medicine, University of Birmingham, Birmingham, United Kingdom.
authors.forename: Gerard B
authors.initials: GB
authors.lastname: Nash
authors.affiliation: School of Chemistry, University of Birmingham, Birmingham, United Kingdom.
authors.forename: James H R
authors.initials: JH
authors.lastname: Tucker
authors.affiliation: Institute of Biomedical Research, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom.
authors.forename: Roy
authors.initials: R
authors.lastname: Bicknell
mesh_terms: D000818:Animals; D016466:CHO Cells; D002232:Carbocyanines / Q000378:metabolism; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D015342:DNA Probes / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism*; D018274:Electroporation; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D008845:Microinjections; D018613:Microscopy, Confocal; D014162:Transfection
chemical_list: D002232:Carbocyanines; D015342:DNA Probes; D004277:DNA, Single-Stranded; C089187:cyanine dye 3; C085321:cyanine dye 5
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