Abstract
SiRNA delivery to the cytoplasm can be obtained through the application of calibrated electric field pulses to a mixture of cells and oligonucleotides. To investigate the uptake pathway, time lapse confocal fluorescence microscopy provides a direct visualization of the transfer. SiRNA is electrophoretically drifted directly to the cytoplasm during the pulse. No post pulse transfer is observed. The uploaded siRNA then freely diffuse in the cytoplasm with no access to the nuclei.
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Acknowledgments
The authors would like to thank laboratory members for their help and comments (Elisabeth Bellard, Aurelie Paganin-Gioanni, Sophie Chabot, Sandrine Pelofy, and Marie-Pierre Rols). Financial supports were obtained from the Seventh Framework European Programme (FP7) OncomiR [grant number 201102], the “Ligue nationale contre le Cancer ,” and the “Région Midi-Pyrenées.”
This work has been performed in collaboration with the “Toulouse Réseau Imagerie” core IPBS facility (Genotoul, Toulouse, France), which is supported by the Association Recherche Cancer (no. 5585), Region Midi Pyrenees (CPER), and Grand Toulouse cluster.
This work was conducted in the scope of the LEA-EBAM and the EU-COST Action TD1104.
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Golzio, M., Teissié, J. (2016). Imaging of Electrotransferred siRNA. In: Medarova, Z. (eds) RNA Imaging. Methods in Molecular Biology, vol 1372. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3148-4_7
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DOI: https://doi.org/10.1007/978-1-4939-3148-4_7
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