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  • Title: Construction of cell penetrating peptide vectors with N-terminal stearylated nuclear localization signal for targeted delivery of DNA into the cell nuclei.
    Author: Wang HY, Chen JX, Sun YX, Deng JZ, Li C, Zhang XZ, Zhuo RX.
    Journal: J Control Release; 2011 Oct 10; 155(1):26-33. PubMed ID: 21187118.
    Abstract:
    Cellular uptake and nuclear localization are two barriers to gene delivery. Here, we designed new gene delivery carriers with an N-terminal stearylated (STR) nuclear localization signal (NLS), PKKKRKV, present in the Simian Virus 40 large T antigen with the aim to overcome limitations, such as cell membrane and nuclear pores, offering attractive possibilities to enhance gene delivery. Four vectors with different structures of N-stearylated nuclear localization signal-octaarginine peptide (STR-PKKKRKV-R(8) or STR-NLS-R(8), STR-VKRKKKP-R(8) or STR-reverse NLS-R(8), PKKKRKV-R(8) or NLS-R(8), and VKRKKKP-R(8) or reverse NLS-R(8)) were compared. The gene expression mediated by these vectors in dividing and non-dividing cells (both in 293T and HeLa cell lines) was investigated. The most efficient N/P ratio was 4 for STR-PKKKRKV-R(8,) STR-VKRKKKP-R(8,) and 0.25 for PKKKRKV-R(8), VKRKKKP-R(8.) The maximum transfection activity of these vehicles (VKRKKKP-R(8)) was up to 80% as effective as jetPEI™ and the vehicles did not exhibit cytotoxicity. Interestingly, N-stearylated peptides presented lower transfection activity compared to peptides without N-stearylation at lower N/P ratios (0.25 to 1). Confocal study showed that the vectors could effectively promote the nuclear translocation.
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