Enhanced plasmid DNA transfection with lysosomotropic agents in cultured fibroblasts

Ciftci, Kadriye; Levy, Robert J.

doi:10.1016/s0378-5173(01)00623-8

Cited by 141 publications

(91 citation statements)

References 28 publications

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“…2). Furthermore, the (L L-Arg) 7 conjugate (PNA2392) showed higher activity than the corresponding D D-Arg conjugate (PNA2785) despite the fact that the D D-peptide should be biologically more stable. In general, …”

Section: Resultsmentioning

confidence: 96%

“…1). In the absence of light, naked PNA showed very little antisense activity in this very sensitive assay system, whereas increased activity was seen for PNA conjugated to oligoarginine ((L L-Arg) 7 , (D D-Arg) 8 , (D D-Arg) 9 and the model amphipathic peptide, KLA (Fig. 1B).…”

Section: Resultsmentioning

confidence: 98%

“…Thus the discovery and development of endosome-disruptive methods are of great interest and several have already been described such as Ca 2+ treatment [5], chloroquine treatment [6], sucrose treatment [7], and photochemical treatment [8]. Among these, photochemical treatment termed photochemical internalization (PCI) may be of particular interest because it has the potential for in vivo applications [9].…”

Section: Introductionmentioning

confidence: 99%

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Photochemically enhanced cellular delivery of cell penetrating peptide‐PNA conjugates

Shiraishi¹,

Nielsen²

2006

FEBS Letters

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Recent studies have shown that endosomal release is a major rate-limiting step for cellular delivery via a variety of cationic cell penetrating peptides. Thus, methods and/or protocols for effective release of endosomally entrapped drugs are highly warranted. Photochemical internalization (PCI) has previously been proposed for this purpose. Here, we demonstrate an enhancement of up to two orders of magnitude of the antisense effects (cytosolic/nuclear) of peptide nucleic acid-peptide conjugates (Tat, Arg 7 , KLA) in HeLa cells by a PCI approach using the photosensitizer AlPc 2a . These results emphasize the importance of endosomal release for cellular activity of this type of drug delivery and also raise hope that methods like PCI which have applications for in vivo use may also enhance the bioavailability and in vivo efficacy of these types of conjugates.

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Section: Resultsmentioning

confidence: 96%

Section: Resultsmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

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Photochemically enhanced cellular delivery of cell penetrating peptide‐PNA conjugates

Shiraishi¹,

Nielsen²

2006

FEBS Letters

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“…At high chloroquine concentrations, endosomes can swell and rupture. [173][174][175] Other chemicals that act via a different mechanism, including sucrose and dimethyl sulfoxide (DMSO), have also been suggested for enhancing PTD mediated delivery. 176,177 Photochemical internalization enhances endosomal release by means of "photosensitizers", which are agents that translocate through intracellular membranes, e.g., some ribonucleases or toxins.…”

Section: ©2 0 1 1 L a N D E S B I O S C I E N C E D O N O T D I S Tmentioning

confidence: 99%

Targeting antibodies to the cytoplasm

Marschall

Frenzel

Schirrmann

et al. 2011

mAbs

110

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“…Besides calciummediated transfection, 46 lysosomotropic agents such as chloroquine were found to improve the transfection of mammalian cells by interfering with several steps involved in transport and release of plasmid DNA from lysosomes. 47 Recent evidence in mice and pigs showed that other tissues may also take up plasmid DNA upon in vivo injection of a supraphysiological dose. 23,48 According to our and others' results, ODN seem to be internalized efficiently into keratinocytes leading to rapid accumulation in the nucleus.…”

Section: Dna-binding Proteins In Keratinocytesmentioning

confidence: 99%

Uptake and trafficking of DNA in keratinocytes: evidence for DNA-binding proteins

et al. 2004

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The skin is an interesting organ for human gene therapy due to accessibility, immunologic potential and synthesis capabilities. In this study, we attempted to visualize and measure the uptake of naked FITC-labeled plasmid by FACS analysis detecting up to 15% internalization in a doseand time-dependent manner. Cycloheximide treatment inhibited the uptake by 490%, suggesting a protein-mediated uptake. The inhibition of different internalization pathways demonstrated that blocking macropinocytosis (by amiloride and N,N-dimethylamylorid) reduced DNA uptake by 485%, while the inhibition of clathrin-coated pits (by chlorpromazine) and caveolae (by nystatin/filipin III) did not limit the uptake. Colocalization studies using confocal laser microscopy revealed a time-dependent accumulation of plasmid DNA in endosomes and lysosomes. When a green fluorescent protein (GFP) expression vector was used, specific GFP-RNA became detectable by reverse transcriptase-PCR, whereas measurable amounts of protein could not be identified in FACS experiments. To detect the potential DNA receptors on the keratinocyte surface, membrane proteins were extracted and subjected to South-Western blotting using digoxigenin-labeled calf thymus and l-phage DNA. Two DNA-binding proteins, ezrin and moesin, known as plasma membrane-actin linkers, were identified by one-and two-dimensional-South-Western blots and matrix-assisted laser desorption and ionizationmass spectrometry. Ezrin and moesin are functionally associated with a number of transmembrane receptors such as the EGF, CD44 or ICAM-1 receptor. Taken together, naked plasmid DNA seems to enter human keratinocytes through different pathways, mainly by macropinocytosis. Two DNA-binding proteins were identified that seemed to be involved in binding/trafficking of internalized DNA.

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Enhanced plasmid DNA transfection with lysosomotropic agents in cultured fibroblasts

Cited by 141 publications

References 28 publications

Photochemically enhanced cellular delivery of cell penetrating peptide‐PNA conjugates

Photochemically enhanced cellular delivery of cell penetrating peptide‐PNA conjugates

Targeting antibodies to the cytoplasm

Uptake and trafficking of DNA in keratinocytes: evidence for DNA-binding proteins

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