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New effects in polynucleotide release from cationic lipid carriers revealed by confocal imaging, fluorescence cross-correlation spectroscopy and single particle tracking

MPG-Autoren
http://pubman.mpdl.mpg.de/cone/persons/resource/persons14835

Berezhna,  S.
Research Group of Experimental Biophysics, MPI for biophysical chemistry, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons15206

Heintzmann,  R.
Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons15268

Jahnz,  M.
Research Group of Experimental Biophysics, MPI for biophysical chemistry, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons14873

Boese,  G.
Research Group of Experimental Biophysics, MPI for biophysical chemistry, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons15815

Schwille,  P.
Research Group of Experimental Biophysics, MPI for biophysical chemistry, Max Planck Society;

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Zitation

Berezhna, S., Schaefer, S., Heintzmann, R., Jahnz, M., Boese, G., Deniz, A., et al. (2005). New effects in polynucleotide release from cationic lipid carriers revealed by confocal imaging, fluorescence cross-correlation spectroscopy and single particle tracking. Biochimica et Biophysica Acta, 1669: doi:10.1016/j.bbamem.2005.02.011, pp. 193-207. Retrieved from http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6T1T-4FNP096-1-C&_cdi=4899&_user=38661&_orig=browse&_coverDate=05%2F20%2F2005&_sk=983309997&view=c&wchp=dGLzVtz-zSkzS&md5=a323cd16432000efe451a74723abc7d2&ie=/sdarticle.pdf.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-0012-E946-0
Zusammenfassung
We report on new insights into the mechanisms of short single and double stranded oligonucleotide release from cationic lipid complexes (lipoplexes), used in gene therapy. Specifically, we modeled endosomal membranes using giant unilamellar vesicles and investigated the roles of various individual cellular phospholipids in interaction with lipoplexes. Our approach uses a combination of confocal imaging, fluorescence cross-correlation spectroscopy and single particle tracking, revealing several new aspects of the release: (a) phosphatidylserine and phosphatidylethanolamine are equally active in disassembling lipoplexes, while phosphatidylcholine and sphingomyelin are inert; (b) in contrast to earlier findings, phosphatidylethanolamine alone, in the absence of anionic phosphatidylserine triggers extensive release; (c) a double-stranded DNA structure remains well preserved after release; (d) lipoplexes exhibited preferential binding to transient lipid domains, which appear at the onset of lipoplex attachment to originally uniform membranes and vanish after initiation of polynucleotide release. The latter effect is likely related to phosphatidyleserine redistribution in membranes due to lipoplex binding. Real time tracking of single DOTAP/DOPE and DOTAP/DOPC lipoplexes showed that both particles remained compact and associated with membranes up to 1–2 min before fusion, indicating that a more complex mechanism, different from suggested earlier rapid fusion, promotes more efficient transfection by DOTAP/DOPE complexes.