Internalization of novel non-viral vector TAT-streptavidin into human cells

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Internalization of novel non-viral vector TAT-streptavidin into human cells

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dc.contributor.author Rinne, Johanna en_US
dc.contributor.author Albarran, Brian en_US
dc.contributor.author Jylhävä, Juulia en_US
dc.contributor.author Ihalainen, Teemu O. en_US
dc.contributor.author Kankaanpää, Pasi en_US
dc.contributor.author Hytönen, Vesa P. en_US
dc.contributor.author Stayton, Patrick S. en_US
dc.contributor.author Kulomaa, Markku S. en_US
dc.contributor.author Vihinen-Ranta, Maija en_US
dc.date.accessioned 2010-05-06T20:06:44Z
dc.date.available 2010-05-06T20:06:44Z
dc.date.issued 2007 en_US
dc.identifier.citation Rinne J, Albarran B, Jylhava J, et al. Internalization of novel non-viral vector TAT-streptavidin into human cells. BMC Biotechnology. 2007;7(1):1. en_US
dc.identifier.other 10.1186/1472-6750-7-1 en_US
dc.identifier.uri http://www.biomedcentral.com/1472-6750/7/1 en_US
dc.identifier.uri http://hdl.handle.net/1773/15850
dc.description.abstract Background: The cell-penetrating peptide derived from the Human immunodeficiency virus-1 transactivator protein Tat possesses the capacity to promote the effective uptake of various cargo molecules across the plasma membrane in vitro and in vivo. The objective of this study was to characterize the uptake and delivery mechanisms of a novel streptavidin fusion construct, TAT[sub]47-57-streptavidin (TAT-SA, 60 kD). SA represents a potentially useful TAT-fusion partner due to its ability to perform as a versatile intracellular delivery vector for a wide array of biotinylated molecules or cargoes. Results: By confocal and immunoelectron microscopy the majority of internalized TAT-SA was shown to accumulate in perinuclear vesicles in both cancer and non-cancer cell lines. The uptake studies in living cells with various fluorescent endocytic markers and inhibiting agents suggested that TAT-SA is internalized into cells efficiently, using both clathrin-mediated endocytosis and lipidraft- mediated macropinocytosis. When endosomal release of TAT-SA was enhanced through the incorporation of a biotinylated, pH-responsive polymer poly(propylacrylic acid) (PPAA), nuclear localization of TAT-SA and TAT-SA bound to biotin was markedly improved. Additionally, no significant cytotoxicity was detected in the TAT-SA constructs. Conclusion: This study demonstrates that TAT-SA-PPAA is a potential non-viral vector to be utilized in protein therapeutics to deliver biotinylated molecules both into cytoplasm and nucleus of human cells. en_US
dc.description.sponsorship Grants from the Academy of Finland (contract 107311) and the NIH (EB00252, DK49655 and CA55596). en_US
dc.language.iso en_US en_US
dc.title Internalization of novel non-viral vector TAT-streptavidin into human cells en_US
dc.type Article en_US


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