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  Characterization of Interaction between Cationic Lipid-Oligonucleotide Complexes and Cellular Membrane Lipids Using Confocal Imaging and Fluorescence Correlation Spectroscopy

Gordon, S. P., Berezhna, S., Scherfeld, D., Kahya, N., & Schwille, P. (2005). Characterization of Interaction between Cationic Lipid-Oligonucleotide Complexes and Cellular Membrane Lipids Using Confocal Imaging and Fluorescence Correlation Spectroscopy. Biophysical Journal, 88(1), 305-316. doi:10.1529/biophysj.104.043133.

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Gordon, S. P., Author
Berezhna, S., Author              
Scherfeld, D., Author              
Kahya, N., Author              
Schwille, P.1, Author              
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1Research Group of Experimental Biophysics, MPI for biophysical chemistry, Max Planck Society, ou_578554              

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 Abstract: Complexes formed by cationic liposomes and single-strand oligodeoxynucleotides (CL-ODN) are promising delivery systems for antisense therapy. ODN release from the complexes is an essential step for inhibiting activity of antisense drugs. We applied fluorescence correlation spectroscopy and confocal laser scanning microscopy to monitor CL-ODN complex interaction with membrane lipids leading to ODN release. To model cellular membranes we used giant unilamellar vesicles and investigated the transport of Cy-5-labeled ODNs across DiO-labeled membranes. For the first time, we directly observed that ODN molecules are transferred across the lipid bilayers and are kept inside the giant unilamellar vesicles after release from the carriers. ODN dissociation from the carrier was assessed by comparing diffusion constants of CL-ODN complexes and ODNs before complexation and after release. Freely diffusing Cy-5-labeled ODN (16-nt) has diffusion constant DODN = 1.3 ± 0.1 × 10−6 cm2/s. Fluorescence correlation spectroscopy curves for CL-ODN complexes were fitted with two components, which both have significantly slower diffusion in the range of DCL-ODN = ∼1.5 × 10−8 cm2/s. Released ODN has the mean diffusion constant D = 1.1 ± 0.2 × 10−6 cm2/s, which signifies that ODN is dissociated from cationic lipids. In contrast to earlier studies, we report that phosphatidylethanolamine can trigger ODN release from the carrier in the full absence of anionic phosphatidylserine in the target membrane and that phosphatidylethanolamine-mediated release is as extensive as in the case of phosphatidylserine. The presented methodology provides an effective tool for probing a delivery potential of newly created lipid formulations of CL-ODN complexes for optimal design of carriers.

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Language(s): eng - English
 Dates: 2005-01
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1529/biophysj.104.043133
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Title: Biophysical Journal
  Other : Biophys. J.
Source Genre: Journal
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Publ. Info: Cambridge, Mass. : Cell Press
Pages: - Volume / Issue: 88 (1) Sequence Number: - Start / End Page: 305 - 316 Identifier: ISSN: 0006-3495
CoNE: https://pure.mpg.de/cone/journals/resource/954925385117