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Título

Efficient Transfection of Hepatocytes Mediated by mRNA Complexed to Galactosylated Cyclodextrins

AutorSymens, Nathalie; Méndez-Ardoy, Alejandro CSIC ORCID; Díaz Moscoso, Alejandro CSIC ORCID ; Sánchez-Fernández, Elena M. CSIC ORCID; Remaut, Katrien; Demeester, Joseph; García Fernández, José Manuel CSIC ORCID; De Smedt, Stefaan; Rejman, Joanna
Fecha de publicación2012
EditorAmerican Chemical Society
CitaciónBioconjugate Chemistry 23: 1276- 1289 (2012)
ResumenIn this study, we aimed at specific targeting of polycationic amphiphilic cyclodextrins (paCDs) to HepG2 cells via the asialoglycoprotein receptor (ASGPr). The transfection efficiencies of paCDs modified with galactose moieties were evaluated. In preliminary experiments, attempts to transfect HepG2 cells with pDNA complexed with different modified paCDs resulted in very low transfection levels. In additional series of experiments, we found out that nucleic acid/cyclodextrin complexes (CDplexes) were efficiently taken up by the cells and that photochemical internalization, which facilitates release from endosomes, did not improve transfection. Further experiments showed that pDNA can be readily released from the CDplexes when exposed to negatively charged vesicles. These observations imply that the lack of transfection cannot be attributed to a lack of internalization, release of CDplexes from the endosomal compartment, or release of free pDNA from the CDplexes. This in turn suggests that the nuclear entry of the pDNA represents the main limiting factor in the transfection process. To verify that HepG2 cells were transfected with targeted CDplexes containing mRNA, which does not require entry into the nucleus for being translated. With mRNA encoding the green fluorescent protein, fractions of GFP-positive cells of up to 31% were obtained. The results confirmed that the galactosylated complexes are specifically internalized via the ASGPr.
URIhttp://hdl.handle.net/10261/99441
DOI10.1021/bc3001003
Identificadoresdoi: 10.1021/bc3001003
issn: 1520-4812
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