Abstract
A series of peptide-based vectors (PolyR8, PolyR8-NLS1, PolyR8-NLS2 and PolyR8-NLS3) containing R8 sequences linked by disulfide bonds are explored as gene vectors. To overcome the nuclear membrane barrier, nuclear localization signal (NLS) sequences were also introduced to these vectors for nuclear import. It was found that all the vectors except PolyR8-NLS3 were efficient for binding and condensing plasmid DNA (pDNA). PolyR8 without an NLS sequence exhibited a relatively high cytotoxicity, while PolyR8-NLS (PolyR8-NLS1, PolyR8-NLS2 and PolyR8-NLS3) displayed low cytotoxicity. Moreover, with the increasing amount of NLS sequences, the cytotoxicity of PolyR8-NLS decreased. PolyR8-NLS2 possessed the highest transfection efficiency at a w/w ratio of 40, which was comparable to jetPEI™. According to the result of flow cytometry, the percentage of cells transfected by PolyR8-NLS2 was 35.7%, which was even superior to that of jetPEI™ (32.0%). PolyR8-NLS2 also presented the best capability for nuclear targeting from the confocal microscopy observations.
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