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RNA Interference Techniques pp 141–157Cite as

Tracing of siRNAs Inside Cells by FRET Imaging

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Part of the book series: Neuromethods ((NM,volume 58))

Abstract

The structural integrity of siRNA carrying fluorescent dyes attached to its different strands can be ­monitored through the efficiency of fluorescence resonance energy transfer between the dyes. In this chapter, the experimental details of the construction of dye-labeled siRNA and of transfection with ­various formulations of siRNAs with cationic lipids, nanoparticles, and liposomes, respectively, are given. Also described are the conditions for confocal microscopy and an algorithm allowing visualization of intact siRNA after image data treatment. The method allows to compare the uptake efficiency mediated by various transfection agents, and to map the location of intact siRNA inside the cell during a timeframe corresponding to a typical knockdown experiment.

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References

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Author information

Authors and Affiliations

  1. Department of Chemistry, Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, Heidelberg, Germany

    Mark Helm

Authors
  1. Markus Hirsch
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  2. Il-Han Kim
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  3. Anne Järve
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  4. Roger Fischer
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  5. Michael F. Trendelenburg
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  6. Ulrich Massing
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  7. Karl Rohr
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  8. Mark Helm
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Corresponding author

Correspondence to Mark Helm .

Editor information

Editors and Affiliations

  1. Research Institute, Center for Gene Therapy, Nationwide Children’s Hospital, Children's Drive 700, Columbus, 43205, Ohio, USA

    Scott Q. Harper

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© 2011 Springer Science+Business Media, LLC

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Hirsch, M. et al. (2011). Tracing of siRNAs Inside Cells by FRET Imaging. In: Harper, S. (eds) RNA Interference Techniques. Neuromethods, vol 58. Humana Press. https://doi.org/10.1007/978-1-61779-114-7_9

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  • DOI: https://doi.org/10.1007/978-1-61779-114-7_9

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-113-0

  • Online ISBN: 978-1-61779-114-7

  • eBook Packages: Springer Protocols

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