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Fluorescence cross‐correlation spectroscopy as a valuable tool to characterize cationic liposome‐DNA nanoparticle assembly
Author(s) -
GómezVarela Ana I.,
Gaspar Ricardo,
Miranda Adelaide,
Assis Juliane L.,
Valverde Rafael H.F.,
EinickerLamas Marcelo,
Silva Bruno F. B.,
De Beule Pieter A.A.
Publication year - 2021
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.202000200
Subject(s) - liposome , cationic liposome , fluorescence correlation spectroscopy , dna , biophysics , chemistry , cationic polymerization , gene delivery , fluorescence , biochemistry , transfection , biology , gene , molecule , organic chemistry , physics , quantum mechanics
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.