Protein transduction: unrestricted delivery into all cells?

Schwarze, Steven R.; Hruska, Keith A.; Dowdy, Steven F.

doi:10.1016/s0962-8924(00)01771-2

Cited by 543 publications

(424 citation statements)

References 39 publications

Supporting

Mentioning

416

Contrasting

Unclassified

Order By: Relevance

“…39 Among these proteins, the HIV protein Tat and the Drosophila Antennapedia homeotic transcription factor have been the most extensively studied. 47 In the case of VP22, it was reported that not only VP22 could enter into cells, but it could be exported from expressing cells. 39 Owing to its intercellular trafficking properties, the use of VP22 has been proposed in gene therapy to translocate therapeutic products from genetically modified cells toward untransduced neighboring cells.…”

Section: Discussionmentioning

confidence: 99%

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

et al. 2004

View full text Add to dashboard Cite

The treatment of solid tumors by retroviral delivery of the herpes simplex virus thymidine kinase (TK) followed by ganciclovir (GCV) treatment has so far shown only limited success in patients. One major drawback in this approach is the lack of efficient in vivo gene delivery to cancer cells. Although, the transduction of every single tumor cell is not a requirement since the bystander effect (BE) mediated by gap junctions allows the diffusion of the toxic GCV metabolites from TK-expressing cells toward untransduced cells. To render the TK/GCV approach more potent, and independent of the level of gap junctions, we have tested the efficiency of a TK mutant (TK30) fused to VP22, a herpes simplex protein that seems to be capable of intercellular trafficking. We failed to detect an increase in the BE with cells expressing VP22 fused to TK30 versus cells containing TK30 alone, and this result forced us to reinvestigate the trafficking properties of VP22. Using very sensitive Western blot and fluorescence assays, we were not able to detect the spread of VP22 fused either to TK30 or GFP. These results indicate that VP22 cannot be used as a cargo to translocate TK30 or GFP.

show abstract

Section: Discussionmentioning

confidence: 99%

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

et al. 2004

View full text Add to dashboard Cite

show abstract

“…108 This function appeared to be imparted by the presence of arginines in the peptides, and could be mimicked by synthetic polyarginine oligopeptides. 109 These basic protein transduction domains (PTDs) have been fused to various cargo to facilitate their cell entry and intracellular trafficking, although the actual mechanism of transduction had been unknown for some time.…”

Section: Intracellular Trafficking Of Nonviral Vectors Lk Medina-kauwmentioning

confidence: 99%

Intracellular trafficking of nonviral vectors

2005

View full text Add to dashboard Cite

“…[1][2][3][4][5][6][7][8][9][10] These membrane-permeable peptides have been used for the delivery of exogenous molecules such as oligonucleotides and proteins into the cytoplasm and the nucleus of cultured cells. Tat-derived peptides that are chemically crosslinked to b-galactosidase, horseradish peroxidase, and RNAase A were found to be successfully taken up by different cell lines.…”

Section: Introductionmentioning

confidence: 99%

Mechanism of improved gene transfer by the N-terminal stearylation of octaarginine: enhanced cellular association by hydrophobic core formation

et al. 2004

View full text Add to dashboard Cite

The internalization mechanisms associated with octaarginine and stearyl-octaarginine were investigated using confocal laser microscopy and flow cytometric analysis. Octaarginine is able to translocate through cell membranes in a manner that does not exactly involve the classical endocytic pathways of internalization. However, when a stearyl moiety is attached to the N-terminus of octaarginine, the internalization shifts mainly to an endocytosis-dependent pathway. The transfection efficiency of stearyl-octaarginine was significantly higher than that of octaarginin. To understand the mechanism of the improved gene transfer by the N-terminal stearylation of octaarginine, the gene transfer processes mediated by octaarginine or stearyl-octaarginine were compared. Both octaarginine and stearyl-octaarginine are able to carry plasmid DNA into cells. The amount of plasmid DNA internalized as well as that delivered to the nucleus was higher in the case of stearyl-octaarginine. Even though the internalization mechanisms of octaarginine and stearyl-octaarginine were different, their complexes with plasmid DNA were internalized via the same pathway, presumably, the clathrin-mediated pathway of endocytosis. The results of the atomic force microscopy revealed that stearyl-octaarginine, but not octaarginine, can completely condense the DNA into stable complexes that can be highly adsorbed to the cell surface and subsequently highly internalized. Therefore, using stearylated-octaarginine provided higher internalization of plasmid DNA into cells, due to enhanced cellular association, as well as higher nuclear delivery. The results presented in this study provide a better understanding of the mechanisms of improved transfection using stearylated-octaarginine. The concept of using stearylated peptides may aid in the development of more efficient nonviral gene vectors.

show abstract

Protein transduction: unrestricted delivery into all cells?

Cited by 543 publications

References 39 publications

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

Intracellular trafficking of nonviral vectors

Mechanism of improved gene transfer by the N-terminal stearylation of octaarginine: enhanced cellular association by hydrophobic core formation

Contact Info

Product

Resources

About