Effect of peptides bearing nuclear localization signals on therapeutic ultrasound mediated gene delivery

Duvshani-Eshet, Maayan; Keren, Hadas; Oz, Shira; Radzishevsky, Inna; Mor, Amram; Machluf, Marcelle

doi:10.1002/jgm.1235

Cited by 22 publications

(19 citation statements)

References 50 publications

(113 reference statements)

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“…The present study confirmed that the NLS and pDNA combined in a complex and the optimum molar ratio for this binding was 10 4 :1 NLS/pDNA, as detected by agarose gel electrophoresis, performed similarly to a previous study by Duvshani-Eshet et al (25). For different exogenous DNAs, the differences in molecular weight and electrostatic charge result in different molar ratios for binding.…”

Section: Discussionsupporting

confidence: 92%

Enhanced effect of nuclear localization signal peptide during ultrasound-targeted microbubble destruction-mediated gene transfection

Cao

Zhou

Chen

et al. 2017

Molecular Medicine Reports

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Ultrasound-targeted microbubble destruction (UTMD) can promote the entry of plasmid DNA (pDNA) into the cell cytoplasm, by increasing the permeability of the cell membrane. But the transfection efficiency remains low due to inability of the pDNA to enter the nucleus. Various methods have been explored to improve the UTMD transfection efficiency, but with little success. In cells, the classic nuclear localization signal (cNLS) peptide is an amino acid sequence that signals proteins that are due for nuclear transport. The present study aimed to investigate whether binding of a cNLS peptide to the pDNA may improve the transfection efficiency of UTMD. Four experimental groups were analyzed: Control group (UTMD + pDNA), group with cNLS (UTMD + pDNA + cNLS), group with mutated NLS (mNLS; UTMD + pDNA + mNLS), and group with cNLS and the nuclear import blocker, wheat germ agglutinin (WGA; UTMD + pDNA + cNLS + WGA). The NLS was labeled by fluorescein isothiocyanate, whereas pDNA was labeled with Cy3. Different molar ratios were tested for the NLS and pDNA combination in order to achieve optimal binding of the two molecules. Human umbilical vein endothelial cells were then transfected using the optimum ultrasonic irradiation parameters and NLS/pDNA molar ratio. At 6 h post-transfection, the rates of Cy3-labeled pDNA inside the cells and their nuclei were detected by flow cytometry and laser confocal microscopy, and the cellular vs. nuclear uptake of pDNA was calculated. In order to further evaluate the effect of NLS on UTMD-mediated gene transfection, the transfection efficiency and relative expression levels of mRNA and protein were detected at 48 h post-transfection. The results demonstrated that the optimal molar ratio of NLS with pDNA was 104:1. The rates of pDNA successful entry into the cell and nucleus were significantly higher in the cNLS group compared with the control group. The transfection efficiency, and relative expression levels of mRNA and protein from the plasmid were significantly increased in the cNLS group compared with the control group. The mNLS group displayed no significant difference compared with the control group, while the WGA group exhibited significant inhibition in most indicators of transfection efficiency compared to the cNLS group. These results suggest that combining a cNLS peptide with pDNA during UTMD-mediated transfection significantly improved transfection efficiency. Thus, a cNLS peptide may be an important mediator and a new strategy in enhancing the efficiency of UTMD-mediated gene transfection.

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Section: Discussionsupporting

confidence: 92%

Enhanced effect of nuclear localization signal peptide during ultrasound-targeted microbubble destruction-mediated gene transfection

Cao

Zhou

Chen

et al. 2017

Molecular Medicine Reports

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“…Therefore, to achieve maximum efficiency CPP, nuclear localized signal peptide, metal NP-liposome based carrier systems have become topics of study. 131 …”

Section: Applications Of Liposome In Drug and Gene Deliverymentioning

confidence: 97%

Liposomes: Versatile and Biocompatible Nanovesicles for Efficient Biomolecules Delivery

Mallick¹,

Choi²

2014

j. nanosci. nanotech.

135

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Since the revolutionary discovery that phospholipids can form closed bilayered structures in aqueous systems, liposomes have become a very interesting topic of research. Because of their versatility and amazing biocompatibility, the use of liposomes has been widely accepted in many scientific disciplines. Their applications, especially in medicine, have yielded breakthroughs with anticancer-drug carriers over the past few decades. Specifically, their easy preparation and various structural aspects have given rise to a broadly usable way to internalize biomolecules such as drugs, DNA, RNA and even imaging probes. This review article reports recent developments in liposomal drug delivery and gene delivery, and thoroughly covers the synthesis and different kinds of liposomal surface modification techniques that have resulted in higher stability and efficiency with respect to the use of liposomes in tumor cell targeting, site-specific release, and extending blood retention times.

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“…15 However, as for NLS peptides alone, other studies have suggested that multidomain peptides, such as a bacterially derived cell-penetrating peptide fused to an NLS, give no increased nuclear localization. 16 Thus, the jury remains out.…”

Section: Nls Peptides Complexed With Plasmids May Enhance Dna Nuclearmentioning

confidence: 99%

Progress and prospects: nuclear import of nonviral vectors

2010

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The nuclear envelope represents a key barrier to successful nonviral transfection and gene therapy both in vitro and in vivo. Although the main purpose of the nuclear envelope is to partition the cell to maintain cytoplasmic components in the cytoplasm and nuclear components, most notably genomic DNA, in the nucleus, this function poses a problem for transfections in which exogenous DNA is delivered into the cytoplasm. After delivery to the cytoplasm, nucleic acids rapidly become complexed with cellular proteins that mediate interactions with the cellular machinery for trafficking. Thus, it is these proteins that, in essence, control the nuclear import of DNA, and we must also understand their activities in cells. In this review, we will discuss the principles of nuclear import of proteins and DNA–protein complexes, as well as the various approaches that investigators have used to improve nuclear targeting of plasmids. These approaches include complexation of plasmids with peptides, native and engineered proteins, ligands and polymers, as well as the inclusion of transcription factor-binding sites for general and cell-specific delivery.

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Effect of peptides bearing nuclear localization signals on therapeutic ultrasound mediated gene delivery

Abstract: This study suggests that CPP-NLS peptides may be used for condensing pDNA and bringing it into the cell cytoplasm, but their ability to mediate nuclear import of pDNA is insignificant.

Cited by 22 publications

References 50 publications

Enhanced effect of nuclear localization signal peptide during ultrasound-targeted microbubble destruction-mediated gene transfection

Enhanced effect of nuclear localization signal peptide during ultrasound-targeted microbubble destruction-mediated gene transfection

Liposomes: Versatile and Biocompatible Nanovesicles for Efficient Biomolecules Delivery

Progress and prospects: nuclear import of nonviral vectors

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