Double-tailed lipid modification as a promising candidate for oligonucleotide delivery in mammalian cells
| Autor: | Ramon Eritja, Begoña Ugarte-Uribe, Santiago Grijalvo, César Martín, Itziar Alkorta, Jon V. Busto, Félix M. Goñi |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
media_common.quotation_subject
Lipid Bilayers Biophysics Oligonucleotides Biochemistry law.invention Model membrane systems Confocal microscopy law Humans Internalization Lipid bilayer Molecular Biology Chromatography High Pressure Liquid media_common DNA Primers Fluorescent Dyes Microscopy Confocal Base Sequence Oligonucleotide Chemistry ACN Transfection Lipids Lipid oligonucleotide conjugates Cellular uptake and localization Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Nucleic acid lipids (amino acids peptides and proteins) Lipid modification Conjugate HeLa Cells |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | Background The potential use of nucleic acids as therapeutic drugs has triggered the quest for oligonucleotide conjugates with enhanced cellular permeability. To this end, the biophysical aspects of previously reported potential lipid oligodeoxyribonucleotide conjugates were studied including its membrane-binding properties and cellular uptake. Methods These conjugates were fully characterized by MALDI-TOF mass spectrometry and HPLC chromatography. Their ability to insert into lipid model membrane systems was evaluated by Langmuir balance and confocal microscopy followed by the study of the internalization of a lipid oligodeoxyribonucleotide conjugate bearing a double-tail lipid modification (C28) into different cell lines by confocal microscopy and flow cytometry. This compound was also compared with other lipid containing conjugates and with the classical lipoplex formulation using Transfectin as transfection reagent. Results This double-tail lipid modification showed better incorporation into both lipid model membranes and cell systems. Indeed, this lipid conjugation was capable of inserting the oligodeoxyribonucleotide into both liquid-disordered and liquid-ordered domains of model lipid bilayer systems and produced an enhancement of oligodeoxyribonucleotide uptake in cells, even better than the effect caused by lipoplexes. In addition, in β2 integrin (CR3) expressing cells this receptor was directly involved in the enhanced internalization of this compound. Conclusions All these features confirm that the dual lipid modification (C28) is an excellent modification for enhancing nucleic acid delivery without altering their binding properties. General significance Compared to the commercial lipoplex approach, oligodeoxyribonucleotide conjugation with C28 dual lipid modification seems to be promising to improve oligonucleotide delivery in mammalian cells. This work was supported with funds from the Spanish Ministry of Economy [Grant BFU2007-62062], the Basque Government [GIV06-42], the Spanish Ministry of Education [Grant CTQ2010-20541], the Generalitat de Catalunya [2009/SGR/208], the Instituto de Salud Carlos III [CB06_01_0019] and Fundación Biofísica Bizkaia. CIBER-BBN is an iniciative funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development. B.U-U. was supported by Universidad de País Vasco-UPV/EHU pre-doctoral fellowship and Fundación Biofísica Bizkaia. J.V.B. was a postdoctoral scientist supported by Universidad de País Vasco-UPV/EHU postdoctoral fellowship. The authors acknowledge the Servicio General de Microscopía Analítica y de Alta Resolución en Biomedicina at the University of Basque Country for assistance with confocal microscopy, Prof. A. Gómez-Muñoz for flow cytometry facilities and Eneritz Bilbao for excellent technical assistance. |
| Databáze: | OpenAIRE |
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