Protein transduction domains (PTDs) are short peptide sequences that can transport wide varieties of cargo across cell membranes. This study assessed the transduction ability of fusion proteins containing optimised variants of the PTD from HIV-1 transactivator of transcription (Tat). Uptake of Tat-PTDs was determined by fluorescent microscopy using the fluorescent protein Venus as a tag, and also by using fusion proteins containing caspase-7 and RhoA bound to Tat-PTD. Upon entering the cytosol the latter two induce apoptosis and the formation of cytoplasmic extensions, morphological changes easily observed by microscopy.
It was found that PTDs with two, three or four sequential Tat-PTD domains could bind to the surface of two of the five cell lines tested. Fluorescent microscopy, however, indicated that the fluorescent constructs remained on the cell surface. As well, PTDs bound to caspase-7 or RhoA did not induce any visible morphological changes in the cells.
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/18794 |
| Date | 12 February 2010 |
| Creators | Komarnicki, Vanessa Adriana Michelle |
| Contributors | Truong, Kevin |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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