title: Construction of cell penetrating peptide vectors with N-terminal stearylated nuclear localization signal for targeted delivery of DNA into the cell nuclei.
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.
authors.affiliation: Key Laboratory of Biomedical Polymers of Ministry of Education & Department of Chemistry, Wuhan University, Wuhan 430072, PR China.
authors.forename: Hui-Yuan
authors.initials: HY
authors.lastname: Wang
authors.forename: Jing-Xiao
authors.initials: JX
authors.lastname: Chen
authors.forename: Yun-Xia
authors.initials: YX
authors.lastname: Sun
authors.forename: Ji-Zhe
authors.initials: JZ
authors.lastname: Deng
authors.forename: Cao
authors.initials: C
authors.lastname: Li
authors.forename: Xian-Zheng
authors.initials: XZ
authors.lastname: Zhang
authors.forename: Ren-Xi
authors.initials: RX
authors.lastname: Zhuo
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D004337:Drug Carriers / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals / Q000737:chemistry*; D009842:Oligopeptides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D013539:Simian virus 40 / Q000737:chemistry*; D014162:Transfection
chemical_list: D004337:Drug Carriers; D019913:Nuclear Localization Signals; D009842:Oligopeptides; D010455:Peptides; C448619:octaarginine; D004247:DNA