Role of endosomes in gene transfection mediated by photochemical internalisation (PCI)

Prasmickaite, Lina; Høgset, Anders; Tjelle, Torunn Elisabeth; Olsen, Vibeke Murberg; Berg, Kristian

doi:10.1002/1521-2254(200011/12)2:6<477::aid-jgm137>3.0.co;2-b

Cited by 69 publications

(58 citation statements)

References 43 publications

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“…It was demonstrated that photochemical treatment, inducing permeabilization of the endocytic vesicles and liberation of the entrapped transfecting genes, can substantially increase the efficiency of gene transfection mediated by nonviral vectors. 24,25 However, in spite of significant increase, they were not able to transfect the entire population of cells compared to the results obtained for photochemically enhanced adenovirus-mediated gene delivery. 26 By relying on the idea that the effectiveness of nonviral transfection systems depends on the cell cycle status, 27 they investigated the role of the cell cycle status in photochemical transfection mediated by two types of synthetic transfection agents: a cationic polypeptide polylysine (polyfection) and a cationic lipid formulation N-(2-amino ethyl)-N,N-dimethyl-2,3-bis(tetradecyloxy)-1-propanaminium bromide/dioleoylphosphatidyl ethanol amine.…”

Section: Intelligent Polymeric Vectors In Gene Therapymentioning

confidence: 83%

Intelligent polymers as nonviral vectors

2005

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The successful gene therapy largely depends on the vector type that allows a selective and efficient gene delivery to target cells with minimal toxicity. Nonviral vectors are much safer and cheaper, can be produced easily in large quantities, and have higher genetic material carrying capacity. However, they are generally less efficient in delivering DNA and initiating gene expression as compared to viral vectors, particularly when used in vivo. As nonviral vectors, polycations may work well for efficient cell uptake and endosomal escape, because they do form compact and smaller complexes with plasmid DNA and carry amine groups, which give positive charge and buffering ability that allows safe escape from endosome/lysosome. However, this is a disadvantage in the following step, which is releasing the plasmid DNA within the cytosol. In order to initiate transcription and enhance gene expression, the polymer/plasmid complex should dissociate after releasing from endosome safely and effectively. There are also other limitations with some of the polycationic carriers, for example, aggregation, toxicity, etc. Intelligent polymers, also called as 'stimuli responsive polymers', have a great potential as nonviral vectors to obtain site-, timing-, and duration period-specific gene expression, which is already exhibited in recent studies that are briefly summarized here. Gene Therapy (2005) 12, S139-S145.

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Section: Intelligent Polymeric Vectors In Gene Therapymentioning

confidence: 83%

Intelligent polymers as nonviral vectors

2005

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“…As described in our earlier work PCI constitutes an efficient and light -inducible method for in vitro transfection with plasmid /polylysine complexes. 3,4,8,9 Unfortunately, plasmid / polylysine complexes are very poor agents for in vivo gene delivery. 10 In contrast, adenoviruses are known to be efficient vectors for in vivo gene delivery.…”

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confidence: 78%

Light-induced adenovirus gene transfer, an efficient and specific gene delivery technology for cancer gene therapy

et al. 2002

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A main issue for further clinical progress of cancer gene therapy is to develop technologies for efficient and specific delivery of therapeutic genes to tumor cells. In this work, we describe a photochemical treatment that substantially improves gene delivery by adenovirus, one of the most efficient gene delivery vectors known. Transduction of two different cell lines was studied by microscopy, flow cytometry, and an enzymatic assay, employing a -galactosidase -encoding adenovirus. The photochemical treatment induced a >20 -fold increase in gene transduction, compared with conventional adenovirus infection, both when measured as the percentage of cells transduced, and when measured as the total -galactosidase activity in the cell population. The effect was most pronounced at lower virus doses, where in some cases the same transduction efficiency could be achieved with a 20 times lower virus dose than with conventional infection. Photochemical treatments are already in clinical use for cancer therapy, and generally are very specific and have few side effects. The photochemical internalization technology described thus has a clear potential for improving both the efficiency and the specificity of gene delivery in cancer gene therapy, making it possible to achieve efficient site -specific in vivo gene delivery by adenoviral vectors.

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“…Interestingly, the photochemical treatment could not counteract the decrease in transgene expression observed at high amounts of polycations. Previous studies suggest that the PCI technology only act intracellularly, 23,24 and thus a possible attenuation of the viral uptake due to aggregate formation would not be counteracted by the photochemical treatment. Some of the decrease in transduction may, however, be attributed to cellular toxicity at high PLL concentrations.…”

Section: Discussionmentioning

confidence: 99%

“…21 The mechanism and benefit of the technology has earlier been documented for nonviral vectors. [22][23][24][25] In this work, the effect of photoactivation of the photosensitizer TPPS 2a (meso-tetraphenylporphine with two sulfonate groups on adjacent phenyl rings) on adenoviral transduction in the presence of the polycationic agents poly-L-lysine (PLL) and SuperFectt is investigated. Our results show that the combined use of polycations and photochemical treatment greatly enhance transduction, especially in cells expressing moderate to low levels of CAR.…”

Section: Introductionmentioning

confidence: 99%