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Fluorescence lifetime imaging microscopy (FLIM) of intracellular transport by means of doubly labelled siRNA architectures

Doll, Larissa 1; Lackner, Jens 2; Rönicke, Franziska 1; Wagenknecht, Hans-Achim 1; Nienhaus, Gerd Ulrich 2,3
1 Institut für Organische Chemie (IOC), Karlsruher Institut für Technologie (KIT)
2 Institut für Angewandte Physik (APH), Karlsruher Institut für Technologie (KIT)
3 Institut für Biologische und Chemische Systeme (IBCS), Karlsruher Institut für Technologie (KIT)

Abstract:

For monitoring the intracellular pathway of small interfering RNA (siRNA), both strands were labelled at internal positions by two ATTO dyes as an interstrand Förster resonance energy transfer pair. siRNA double strands show red emission and a short donor lifetime as readout, whereas siRNA antisense single strands show green emission and a long donor lifetime. This readout signals if GFP silencing can be expected (green) or not (red). We attached both dyes to three structurally different alkyne anchors by postsynthetic modifications. There is only a slight preference for the ribofuranoside anchors with the dyes at their 2’-positions. For the first time, the delivery and fate of siRNA in live HeLa cells was tracked by fluorescence lifetime imaging microscopy (FLIM), which revealed a clear relationship between intracellular transport using different transfection methods and knockdown of GFP expression, which demonstrates the potential of our siRNA architectures as a tool for future development of effective siRNA.


Verlagsausgabe §
DOI: 10.5445/IR/1000134221
Veröffentlicht am 26.04.2022
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Angewandte Physik (APH)
Institut für Biologische und Chemische Systeme (IBCS)
Institut für Nanotechnologie (INT)
Institut für Organische Chemie (IOC)
Publikationstyp Zeitschriftenaufsatz
Publikationsdatum 03.08.2021
Sprache Englisch
Identifikator ISSN: 1439-4227, 1439-7633
KITopen-ID: 1000134221
HGF-Programm 43.32.03 (POF IV, LK 01) Designed Optical Devices & Systems
Erschienen in ChemBioChem
Verlag Wiley-VCH Verlag
Band 22
Heft 15
Seiten 2561-2567
Vorab online veröffentlicht am 14.06.2021
Nachgewiesen in Dimensions
Scopus
Web of Science
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