Fluorescence cross-correlation spectroscopy as a valuable tool to characterize cationic liposome-DNA nanoparticle assembly.

Autor: Gómez-Varela AI; International Iberian Nanotechnology Laboratory, Braga, Portugal.; Department of Applied Physics, University of Santiago de Compostela, Santiago de Compostela, Spain., Gaspar R; International Iberian Nanotechnology Laboratory, Braga, Portugal., Miranda A; International Iberian Nanotechnology Laboratory, Braga, Portugal., Assis JL; Biomembranes Laboratory, Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil., Valverde RHF; Biomembranes Laboratory, Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil., Einicker-Lamas M; Biomembranes Laboratory, Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil., Silva BFB; International Iberian Nanotechnology Laboratory, Braga, Portugal., De Beule PAA; International Iberian Nanotechnology Laboratory, Braga, Portugal.
Jazyk: angličtina
Zdroj: Journal of biophotonics [J Biophotonics] 2021 Jan; Vol. 14 (1), pp. e202000200. Date of Electronic Publication: 2020 Oct 01.
DOI: 10.1002/jbio.202000200
Abstrakt: The development of nonviral gene delivery vehicles for therapeutic applications requires methods capable of quantifying the association between the genes and their carrier counterparts. Here we investigate the potential of fluorescence cross-correlation spectroscopy (FCCS) to characterize and optimize the assembly of nonviral cationic liposome (CL)-DNA complexes based on a CL formulation consisting of the cationic lipid DOTAP and zwitterionic lipid DOPC. We use a DNA plasmid for lipoplex loading encoding the Oct4 gene, critically involved in reprogramming somatic cells into induced pluripotent stem cells. We demonstrate that FCCS is able to quantitatively determine the extent of the association between DNA and the liposomes and assess its loading capacity. We also establish that the cationic lipid fraction, being proportional to the liposome membrane charge density, as well as charge ratio between the CLs and anionic DNA play an important role in the degree of interaction between the liposomes and DNA.
(© 2020 Wiley-VCH GmbH.)
Databáze: MEDLINE
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