Dye selection for live cell imaging of intact siRNA
- Publikationstyp:
- Zeitschriftenaufsatz
- Metadaten:
-
- Autoren
- Markus Hirsch
- Dennis Strand
- Mark Helm
- Autoren-URL
- https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000300741400003&DestLinkType=FullRecord&DestApp=WOS_CPL
- DOI
- 10.1515/BC-2011-256
- eISSN
- 1437-4315
- Externe Identifier
- Clarivate Analytics Document Solution ID: 898KN
- PubMed Identifier: 22628296
- ISSN
- 1431-6730
- Ausgabe der Veröffentlichung
- 1-2
- Zeitschrift
- BIOLOGICAL CHEMISTRY
- Schlüsselwörter
- confocal fluorescence microscopy
- fluorescence resonance energy transfer (FRET)
- RNA interference (RNAi)
- siRNA degradation
- siRNA integrity
- small interfering RNA (siRNA)
- Paginierung
- 23 - 35
- Datum der Veröffentlichung
- 2012
- Status
- Published
- Titel
- Dye selection for live cell imaging of intact siRNA
- Sub types
- Article
- Ausgabe der Zeitschrift
- 393
Datenquelle: Web of Science (Lite)
- Andere Metadatenquellen:
-
- Abstract
- <jats:title>Abstract</jats:title> <jats:p>Investigations into the fate of small interfering RNA (siRNA) after transfection may unravel new ways to improve RNA interference (RNAi) efficiency. Because intracellular degradation of RNA may prevent reliable observation of fluorescence-labeled siRNA, new tools for fluorescence microscopy are warranted to cover the considerable duration of the RNAi effect. Here, the characterization and application of new fluorescence resonance energy transfer (FRET) dye pairs for sensing the integrity of duplex siRNA is reported, which allows an assessment of the degradation status of an siRNA cell population by live cell imaging. A panel of high-yield fluorescent dyes has been investigated for their suitability as FRET pairs for the investigation of RNA inside the cell. Nine dyes in 13 FRET pairs were evaluated based on the performance in assays of photostability, cross-excitation, bleed-through, as well as on quantified changes of fluorescence as a consequence of, e.g., RNA strand hybridization and pH variation. The Atto488/Atto590 FRET pair has been applied to live cell imaging, and has revealed first aspects of unusual trafficking of intact siRNA. A time-lapse study showed highly dynamic movement of siRNA in large perinuclear structures. These and the resulting optimized FRET labeled siRNA are expected to have significant impact on future observations of labeled RNAs in living cells.</jats:p>
- Autoren
- Markus Hirsch
- Dennis Strand
- Mark Helm
- DOI
- 10.1515/bc-2011-256
- eISSN
- 1437-4315
- ISSN
- 1431-6730
- Ausgabe der Veröffentlichung
- 1-2
- Zeitschrift
- Biological Chemistry
- Paginierung
- 23 - 35
- Datum der Veröffentlichung
- 2012
- Status
- Published
- Herausgeber
- Walter de Gruyter GmbH
- Herausgeber URL
- http://dx.doi.org/10.1515/bc-2011-256
- Datum der Datenerfassung
- 2021
- Titel
- Dye selection for live cell imaging of intact siRNA
- Ausgabe der Zeitschrift
- 393
Datenquelle: Crossref
- Abstract
- Investigations into the fate of small interfering RNA (siRNA) after transfection may unravel new ways to improve RNA interference (RNAi) efficiency. Because intracellular degradation of RNA may prevent reliable observation of fluorescence-labeled siRNA, new tools for fluorescence microscopy are warranted to cover the considerable duration of the RNAi effect. Here, the characterization and application of new fluorescence resonance energy transfer (FRET) dye pairs for sensing the integrity of duplex siRNA is reported, which allows an assessment of the degradation status of an siRNA cell population by live cell imaging. A panel of high-yield fluorescent dyes has been investigated for their suitability as FRET pairs for the investigation of RNA inside the cell. Nine dyes in 13 FRET pairs were evaluated based on the performance in assays of photostability, cross-excitation, bleed-through, as well as on quantified changes of fluorescence as a consequence of, e.g., RNA strand hybridization and pH variation. The Atto488/Atto590 FRET pair has been applied to live cell imaging, and has revealed first aspects of unusual trafficking of intact siRNA. A time-lapse study showed highly dynamic movement of siRNA in large perinuclear structures. These and the resulting optimized FRET labeled siRNA are expected to have significant impact on future observations of labeled RNAs in living cells.
- Addresses
- Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Im Neuenheimer Feld 364, D-69120 Heidelberg, Germany.
- Autoren
- Markus Hirsch
- Dennis Strand
- Mark Helm
- DOI
- 10.1515/bc-2011-256
- eISSN
- 1437-4315
- Externe Identifier
- PubMed Identifier: 22628296
- Open access
- false
- ISSN
- 1431-6730
- Ausgabe der Veröffentlichung
- 1-2
- Zeitschrift
- Biological chemistry
- Schlüsselwörter
- Brain
- Cells, Cultured
- Endothelial Cells
- Animals
- Rats
- RNA, Small Interfering
- Fluorescent Dyes
- Microscopy, Confocal
- Fluorescence Resonance Energy Transfer
- Cell Survival
- Hydrogen-Ion Concentration
- Sprache
- eng
- Medium
- Paginierung
- 23 - 35
- Datum der Veröffentlichung
- 2012
- Status
- Published
- Datum der Datenerfassung
- 2012
- Titel
- Dye selection for live cell imaging of intact siRNA.
- Sub types
- Research Support, Non-U.S. Gov't
- Journal Article
- Ausgabe der Zeitschrift
- 393
Datenquelle: Europe PubMed Central
- Abstract
- Investigations into the fate of small interfering RNA (siRNA) after transfection may unravel new ways to improve RNA interference (RNAi) efficiency. Because intracellular degradation of RNA may prevent reliable observation of fluorescence-labeled siRNA, new tools for fluorescence microscopy are warranted to cover the considerable duration of the RNAi effect. Here, the characterization and application of new fluorescence resonance energy transfer (FRET) dye pairs for sensing the integrity of duplex siRNA is reported, which allows an assessment of the degradation status of an siRNA cell population by live cell imaging. A panel of high-yield fluorescent dyes has been investigated for their suitability as FRET pairs for the investigation of RNA inside the cell. Nine dyes in 13 FRET pairs were evaluated based on the performance in assays of photostability, cross-excitation, bleed-through, as well as on quantified changes of fluorescence as a consequence of, e.g., RNA strand hybridization and pH variation. The Atto488/Atto590 FRET pair has been applied to live cell imaging, and has revealed first aspects of unusual trafficking of intact siRNA. A time-lapse study showed highly dynamic movement of siRNA in large perinuclear structures. These and the resulting optimized FRET labeled siRNA are expected to have significant impact on future observations of labeled RNAs in living cells.
- Date of acceptance
- 2011
- Autoren
- Markus Hirsch
- Dennis Strand
- Mark Helm
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/22628296
- DOI
- 10.1515/BC-2011-256
- eISSN
- 1437-4315
- Ausgabe der Veröffentlichung
- 1-2
- Zeitschrift
- Biol Chem
- Schlüsselwörter
- Animals
- Brain
- Cell Survival
- Cells, Cultured
- Endothelial Cells
- Fluorescence Resonance Energy Transfer
- Fluorescent Dyes
- Hydrogen-Ion Concentration
- Microscopy, Confocal
- RNA, Small Interfering
- Rats
- Sprache
- eng
- Country
- Germany
- Paginierung
- 23 - 35
- PII
- /j/bchm.2012.393.issue-1-2/bc-2011-256/bc-2011-256.xml
- Datum der Veröffentlichung
- 2012
- Status
- Published
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2013
- Titel
- Dye selection for live cell imaging of intact siRNA.
- Sub types
- Journal Article
- Research Support, Non-U.S. Gov't
- Ausgabe der Zeitschrift
- 393
Datenquelle: PubMed
- Beziehungen:
- Eigentum von