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Evaluation of Different Photosensitizers for Use in Photochemical Gene Transfection ¶
Author(s) -
Prasmickaite Lina,
Høgset Anders,
Berg Kristian
Publication year - 2001
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1562/0031-8655(2001)0730388eodpfu2.0.co2
Subject(s) - transfection , photosensitizer , chemistry , endocytosis , polylysine , gene delivery , endocytic cycle , biophysics , photodynamic therapy , vesicle , endosome , microbiology and biotechnology , biochemistry , photochemistry , intracellular , biology , gene , membrane , cell , organic chemistry
Many potentially therapeutic macromolecules, e.g. transgenes used in gene therapy, are taken into the cells by endocytosis, and have to be liberated from endocytic vesicles in order to express a therapeutic function. To achieve this we have developed a new technology, named photochemical internalization (PCI), based on photochemical reactions inducing rupture of endocytic vesicles. The aim of this study was to clarify which properties of photosensitizers are important for obtaining the PCI effect improving gene transfection. The photochemical effect on transfection of human melanoma THX cells has been studied employing photosensitizers with different physicochemical properties and using two gene delivery vectors: the cationic polypeptide polylysine and the cationic lipid 1,2‐dioleoyl‐3‐trimethylammonium‐propane (DOTAP). Photochemical treatment by photosensitizers that do not localize in endocytic vesicles (tetra[3‐hydroxyphenyl]porphyrin and 5‐aminolevulinic acid–induced protoporphyrin IX) do not stimulate transfection, irrespective of the gene delivery vector. In contrast, photosensitizers localized in endocytic vesicles stimulate polylysine‐mediated transfection, and amphiphilic photosensitizers (disulfonated aluminium phthalocyanine [AlPcS 2a ] and meso ‐tetraphenylporphynes) show the strongest positive effect, inducing approximately 10‐fold increase in transfection efficiency. In contrast, DOTAP‐mediated transfection is inhibited by all photochemical treatments irrespective of the photosensitizer used. Neither AlPcS 2a nor Photofrin affects the uptake of the transfecting DNA over the plasma membrane, therefore photochemical permeabilization of endocytic vesicles seems to be the most likely mechanism responsible for the positive PCI effect on gene transfection.