Construction of cell penetrating peptide vectors with N-terminal stearylated nuclear localization signal for targeted delivery of DNA into the cell nuclei

Wang, Huiyuan; Chen, Jingxiao; Sun, Yunxia; Deng, Jizhe; Li, Cao; Zhang, Xian‐Zheng; Zhuo, Ren‐Xi

doi:10.1016/j.jconrel.2010.12.009

Cited by 126 publications

(82 citation statements)

References 28 publications

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“…17 A single mutation of a lysine residue to a proline residue (PKPKRKV) in NLS limits its nuclear translocation but enhances release of the cargo into the cytoplasm, which is helpful for siRNA intracellular localization. 18 Recently, stearylation has proven to be successful in increasing the transfection efficiency of CPPs for DNA delivery [19][20][21][22][23][24][25] or that of polyethyleneimine for siRNA delivery. 26 The increased transfection efficiency is attributed to the enhanced compaction of DNA, increased endosomal escape, and higher cellular uptake, which are all conferred by the stearyl moiety.…”

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confidence: 99%

Effective small interfering RNA delivery in vitro via a new stearylated cationic peptide

Chen¹,

Ran²,

Askhatova³

et al. 2015

IJN

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A crucial bottleneck in RNA interference-based gene therapy is the lack of safe and efficient delivery systems. Here, a novel small interfering RNA (siRNA) delivery peptide, STR-HK, was constructed by conjugating a stearyl end to the N-terminus of the peptide sequence HHHPKPKRKV, where PKPKRKV is an altered sequence of the nucleus localization signal (PKKKRKV) and contributes to the cytosol localization of STR-HK–siRNA complexes. Histidine is a linker and plays an important role in disrupting the endosomal membrane via the proton sponge effect. As expected, STR-HK formed complexes with siRNA with a particle size of 80–160 nm in diameter and efficiently delivered Cy3-labeled glyceraldehyde 3-phosphate dehydrogenase siRNA into PC-3 human prostate cancer cells. The transfection efficiency of STR-HK at molar ratio of 60/1 was comparable to that of Lipofectamine 2000, one of the most efficient commercially available transfection reagents. Furthermore, the STR-HK–siRNA complexes exhibited minimal cytotoxicity, which was significantly lower than that of Lipofectamine. Taken together, the strategy of conjugating the stearyl moiety with HHHPKPKRKV as a non-viral siRNA delivery system is advantageous.

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confidence: 99%

Effective small interfering RNA delivery in vitro via a new stearylated cationic peptide

Chen¹,

Ran²,

Askhatova³

et al. 2015

IJN

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“…As a result, NLSs should not be used in isolation but rather as supplements to existing vectors. For example functionalization of R8 with the SV40 NLS provoked transfection efficiency that was almost as effective (80%) as jetPEI TM (transfection reagent), but lacked jetPEI TM 's cytotoxicity in HeLa cells [115].…”

Section: Facilitating Nuclear Importmentioning

confidence: 99%

Delivery of Nucleic Acids for Cancer Gene therapy: Overcoming extra- and intra-cellular Barriers

2016

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The therapeutic potential of cancer gene therapy has been limited by the difficulty of delivering genetic material to target sites. Various biological and molecular barriers exist which need to be overcome before effective non-viral delivery systems can be applied successfully in oncology.Herein, various barriers are described and strategies to circumvent such obstacles are discussed, considering both the extracellular and intracellular setting. Development of multifunctional delivery systems holds much promise for the progression of gene delivery, and a growing body of evidence supports this approach involving rational design of vectors, with a unique molecular architecture. In addition, the potential application of composite gene delivery platforms is highlighted which may provide an alternative delivery strategy to traditional systemic administration.

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“…16,17 The use of NLSs to promote nuclear importation and enhance gene delivery has been reported in various non-viral gene delivery vectors including cationic polymers, [18][19][20] oligopeptides 21 and cell penetrating peptides. 22,23 Previously, our group and others have reported the use of LAH4-L1 peptide for DNA and siRNA delivery. [24][25][26][27][28] LAH4-L1 is a histidine-rich, cationic amphipathic peptide with pH responsive properties.…”

Section: Introductionmentioning

confidence: 99%

Incorporation of a Nuclear Localization Signal in pH Responsive LAH4-L1 Peptide Enhances Transfection and Nuclear Uptake of Plasmid DNA

Liang

Qiu

et al. 2016

Mol. Pharmaceutics

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The major intracellular barriers associated with DNA delivery using non-viral vectors are inefficient endosomal/lysosomal escape and poor nuclear uptake. LAH4-L1, a pH responsive cationic amphipathic peptide, is an efficient DNA delivery vector that promotes the release of nucleic acid into cytoplasm through endosomal escape. Here we further enhance the DNA transfection efficiency of LAH4-L1 by incorporating nuclear localizing signal (NLS) to promote nuclear importation. Four NLSs were investigated: Simian virus 40 (SV40) large T-antigen derived NLS, nucleoplasmin targeting signal, M9 sequence and the reverse SV40 derived NLS.All peptides tested were able to form positively charged nano-sized complexes with DNA.Significant improvement in DNA transfection was observed in slow-dividing epithelial cancer cells (Calu-3), macrophages (RAW264.7), dendritic cells (JAWSII), as well as thymidine-induced growth-arrested cells, but not in rapidly dividing cells (A549). Among the four NLS-modified peptides, PK1 (modified with SV40 derived NLS) and PK2 (modified with reverse SV40 derived NLS) were the most consistent in improving DNA transfection; up to a 10-fold increase in gene expression was observed for PK1 and PK2 over the unmodified LAH4-L1. Additionally PK1 and PK2 were shown to enhance cellular uptake as well as nuclear entry of DNA. Overall, we show that the incorporation of SV40 derived NLS, in particular, to LAH4-L1 is a promising strategy to improve DNA delivery efficiency in slow-dividing cells and dendritic cells, with development potential for in vivo applications and as a DNA vaccine carrier.

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Construction of cell penetrating peptide vectors with N-terminal stearylated nuclear localization signal for targeted delivery of DNA into the cell nuclei

Cited by 126 publications

References 28 publications

Effective small interfering RNA delivery in vitro via a new stearylated cationic peptide

Effective small interfering RNA delivery in vitro via a new stearylated cationic peptide

Delivery of Nucleic Acids for Cancer Gene therapy: Overcoming extra- and intra-cellular Barriers

Incorporation of a Nuclear Localization Signal in pH Responsive LAH4-L1 Peptide Enhances Transfection and Nuclear Uptake of Plasmid DNA

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