title: FRET imaging of cells transfected with siRNA/liposome complexes.
abstract: By monitoring the efficiency of fluorescence resonance energy transfer of dyes attached to the different strands of siRNA, the structural integrity of the latter can be traced inside cells. Here, the experimental details of dye-labeled siRNA construction, tissue culture, and transfection with liposomally formulated siRNAs are given, as well as the conditions for confocal microscopy and an algorithm allowing the visualization of intact siRNA after image data treatment. The method allows rapid screening of different liposomal siRNA formulations, obtained by small scale dual asymmetric centrifugation with high entrapping efficiency.
authors.affiliation: Department of Bioinformatics and Functional Genomics, German Cancer Research Center, Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, Germany.
authors.forename: Il-Han
authors.initials: IH
authors.lastname: Kim
authors.forename: Anne
authors.initials: A
authors.lastname: Järve
authors.forename: Markus
authors.initials: M
authors.lastname: Hirsch
authors.forename: Roger
authors.initials: R
authors.lastname: Fischer
authors.forename: Michael F
authors.initials: MF
authors.lastname: Trendelenburg
authors.forename: Ulrich
authors.initials: U
authors.lastname: Massing
authors.forename: Karl
authors.initials: K
authors.lastname: Rohr
authors.forename: Mark
authors.initials: M
authors.lastname: Helm
mesh_terms: D000818:Animals; D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology; D031541:Fluorescence Resonance Energy Transfer / Q000191:economics / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis; D006801:Humans; D008081:Liposomes / Q000737:chemistry; D008845:Microinjections; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000032:analysis*; D051381:Rats; D016707:Tissue Fixation; D014162:Transfection
chemical_list: D005456:Fluorescent Dyes; D008081:Liposomes; D034741:RNA, Small Interfering