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Title

Biophysics and protein corona analysis of Janus cyclodextrin-DNA nanocomplexes. Efficient cellular transfection on cancer cells

AuthorsMartínez-Negro, M.; Caracciolo, G.; Palchetti, S.; Pozzi, D.; Capriotti, A.L.; Cavaliere, C.; Laganà, A.; Ortiz-Mellet, Carmen; Benito, Juan M. ; García-Fernández, José Manuel; Aicart, Emilio
KeywordsCellular transfection
Effective charge ratio
Cytotoxicity
Protein corona
Multilamellar phases
CDplexes
Issue Date2017
PublisherElsevier
CitationBiochimica et Biophysica Acta - General Subjects 1861: 1737- 1749 (2017)
AbstractThe self-assembling processes underlining the capabilities of facially differentiated (¿Janus¿) polycationic amphiphilic cyclodextrins (paCDs) as non-viral gene nanocarriers have been investigated by a pluridisciplinary approach. Three representative Janus paCDs bearing a common tetradecahexanoyl multitail domain at the secondary face and differing in the topology of the cluster of amino groups at the primary side were selected for this study. All of them compact pEGFP-C3 plasmid DNA and promote transfection in HeLa and MCF-7 cells, both in absence and in presence of human serum. The electrochemical and structural characteristics of the paCD-pDNA complexes (CDplexes) have been studied by using zeta potential, DLS, SAXS, and cryo-TEM. paCDs and pDNA, when assembled in CDplexes, render effective charges that are lower than the nominal ones. The CDplexes show a self-assembling pattern corresponding to multilamellar lyotropic liquid crystal phases, characterized by a lamellar stacking of bilayers of the CD-based vectors with anionic pDNA sandwiched among them. When exposed to human serum, either in the absence or in the presence of pDNA, the surface of the cationic CD-based vector becomes coated by a protein corona (PC) whose composition has been analyzed by nanoLC-MS/MS. Some of the CDplexes herein studied showed moderate-to-high transfection levels in HeLa and MCF-7 cancer cells combined with moderate-to-high cell viabilities, as determined by FACS and MTT reduction assays. The ensemble of data provides a detail picture of the paCD-pDNA-PC association processes and a rational base to exploit the protein corona for targeted gene delivery on future in vivo applications.
URIhttp://hdl.handle.net/10261/156158
DOI10.1016/j.bbagen.2017.03.010
Identifiersdoi: 10.1016/j.bbagen.2017.03.010
issn: 1872-8006
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