Fluorescence Lifetime Imaging Microscopy (FLIM) of Intracellular Transport by Means of Doubly Labelled siRNA Architectures.
cycloaddition
energy transfer
fluorescence
oligonucleotide
transport
Journal
Chembiochem : a European journal of chemical biology
ISSN: 1439-7633
Titre abrégé: Chembiochem
Pays: Germany
ID NLM: 100937360
Informations de publication
Date de publication:
03 08 2021
03 08 2021
Historique:
revised:
02
06
2021
received:
30
03
2021
pubmed:
15
6
2021
medline:
20
1
2022
entrez:
14
6
2021
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 34125482
doi: 10.1002/cbic.202100150
pmc: PMC8453559
doi:
Substances chimiques
RNA, Small Interfering
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2561-2567Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : 2039/2
Organisme : Helmholtz Association
Informations de copyright
© 2021 The Authors. ChemBioChem published by Wiley-VCH GmbH.
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