scholarly journals Arginine-based cationic liposomes for efficient in vitro plasmid DNA delivery with low cytotoxicity

2013 ◽  
pp. 1361 ◽  
Author(s):  
Shinji Takeoka ◽  
Sarker ◽  
Aoshima ◽  
Hokama ◽  
Inoue ◽  
...  
Keyword(s):  
Plasmid Dna ◽  
Dna Delivery ◽  
Cationic Liposomes ◽  
Low Cytotoxicity

Evaluation of biocompatibility and efficiency of plasmid DNA delivery by gene-activated hydrogels in vitro

2019 ◽  
Vol XIV (2) ◽  
Author(s):  
I.Y. Bozo ◽  
A.A. Titova ◽  
M.N. Zhuravleva ◽  
A.I. Bilyalov ◽  
M.O. Mavlikeev ◽  
...  
Keyword(s):  
Plasmid Dna ◽  
Dna Delivery

scholarly journals In Vitro Gene Delivery Mediated by Asialofetuin-Appended Cationic Liposomes Associated with γ-Cyclodextrin into Hepatocytes

2011 ◽  
Vol 2011 ◽  
pp. 1-13 ◽  
Author(s):  
Keiichi Motoyama ◽  
Yoshihiro Nakashima ◽  
Yukihiko Aramaki ◽  
Fumitoshi Hirayama ◽  
Kaneto Uekama ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Gene Delivery ◽  
Plasmid Dna ◽  
Transfection Efficiency ◽  
Cationic Liposomes ◽  
Nonviral Vector ◽  
Liposomal Membrane ◽  
Transfer Activity ◽  
Potential Use

The purpose of this study is to evaluate in vitro gene delivery mediated by asialofetuin-appended cationic liposomes (AF-liposomes) associating cyclodextrins (CyD/AF-liposomes) as a hepatocyte-selective nonviral vector. Of various CyDs, AF-liposomes associated with plasmid DNA (pDNA) and γ-cyclodextrin (γ-CyD) (pDNA/γ-CyD/AF-liposomes) showed the highest gene transfer activity in HepG2 cells without any significant cytotoxicity. In addition, γ-CyD enhanced the encapsulation ratio of pDNA with AF-liposomes, and also increased gene transfer activity as the entrapment ratio of pDNA into AF-liposomes was increased. γ-CyD stabilized the liposomal membrane of AF-liposomes and inhibited the release of calcein from AF-liposomes. The stabilizing effect of γ-CyD may be, at least in part, involved in the enhancing gene transfer activity of pDNA/γ-CyD/AF-liposomes. Therefore, these results suggest the potential use of γ-CyD for an enhancer of transfection efficiency of AF-liposomes.

scholarly journals TAMI-14. NANOPARTICLE DELIVERY OF PD-L1 CRISPR/CAS9 PLASMID DNA FOR ANTI-GLIOBLASTOMA IMMUNOTHERAPY

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii216-ii216
Author(s):  
Javier Fierro ◽  
An Tran ◽  
Chris Factoriza ◽  
Brandon Chin ◽  
Huanyu Dou
Keyword(s):  
Plasmid Dna ◽  
Viral Vectors ◽  
New Technologies ◽  
Transfection Efficiency ◽  
Guide Rna ◽  
Immune Balance ◽  
Main Challenge ◽  
Low Cytotoxicity

Abstract Glioblastoma multiforme (GBM) is a devastating cancer that develops from astrocytes in the brain. GBM is fast acting and kills 90% of patients within 5 years. Several immunotherapies have been developed to treat GBM, however, major challenges still persist. For example, checkpoint proteins such as programmed cell death protein 1 (PD-1) and its ligand, programmed death ligand 1 (PD-L1), are upregulated in GBM cells to evade the immune system. Targeting PD-L1 for genetic knockdown is thus a promising avenue for the treatment of GBM. However, PD-L1 protein inhibitors have been shown to cause immune overreaction and toxicity, therefore requiring new technologies. CRISPR/Cas9 gene editing has been widely used for the study and treatment of many diseases, but has not been extensively studied for the treatment of GBM. The main challenge is developing a gene delivery platform for the delivery of CRISPR/Cas9 plasmid DNA (pDNA). Many viral vectors have been used for the delivery of pDNA, but unfortunately are associated with high toxicity. Nanotechnology is emerging as a new platform for the delivery of pDNA as it shows high transfection efficiency with low cytotoxicity. We developed a cationic core-shell nanoparticle (NP) capable of carrying CRISPR/Cas9 pDNA. This plasmid contains multiple guide RNA (gRNA) expression cassettes for the knockdown of PD-L1. PDL1gRNA-CRISPR/Cas9pDNA-NPs were taken up by U87 cells within 30 minutes, and entered into the nucleus at 2 hours. The effective delivery of PDL1gRNA-CRISPR/Cas9pDNA-NPs led to the expression of PD-L1 gRNA and Cas9 enzyme, and the knockdown of PD-L1. Regulation of immune balance was determined after PD-L1 knockdown in vitro and in vivo. Our study shows the potential of NP-based PDL1gRNA-CRISPR/Cas9 delivery as an anti-GBM immunotherapy for clinical applications.

scholarly journals Citrate-Coated Magnetic Polyethyleneimine Composites for Plasmid DNA Delivery into Glioblastoma

Polymers ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 2228
Author(s):  
Ken Cham-Fai Leung ◽  
Kathy W. Y. Sham ◽  
Josie M. Y. Lai ◽  
Yi-Xiang J. Wang ◽  
Chi-Hin Wong ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Plasmid Dna ◽  
Green Fluorescence Protein ◽  
Superparamagnetic Iron Oxide ◽  
Blue Staining ◽  
Luciferase Expression ◽  
Ultrasmall Superparamagnetic Iron Oxide ◽  
Low Cytotoxicity ◽  
Optimal Efficiency

Several ternary composites that are based on branched polyethyleneimine (bPEI 25 kDa, polydispersity 2.5, 0.1 or 0.2 ng), citrate-coated ultrasmall superparamagnetic iron oxide nanoparticles (citrate-NPs, 8–10 nm, 0.1, 1.0, or 2.5 µg), and reporter circular plasmid DNA pEGFP-C1 or pRL-CMV (pDNA 0.5 µg) were studied for optimization of the best composite for transfection into glioblastoma U87MG or U138MG cells. The efficiency in terms of citrate-NP and plasmid DNA gene delivery with the ternary composites could be altered by tuning the bPEI/citrate-NP ratios in the polymer composites, which were characterized by Prussian blue staining, in vitro magnetic resonance imaging as well as green fluorescence protein and luciferase expression. Among the composites prepared, 0.2 ng bPEI/0.5 μg pDNA/1.0 µg citrate-NP ternary composite possessed the best cellular uptake efficiency. Composite comprising 0.1 ng bPEI/0.5 μg pDNA/0.1 μg citrate-NP gave the optimal efficiency for the cellular uptake of the two plasmid DNAs to the nucleus. The best working bPEI concentration range should not exceed 0.2 ng/well to achieve a relatively low cytotoxicity.

scholarly journals Physicochemical Factors That Influence the Biocompatibility of Cationic Liposomes and Their Ability to Deliver DNA to the Nuclei of Ovarian Cancer SK-OV-3 Cells

Materials ◽  
2021 ◽  
Vol 14 (2) ◽  
pp. 416 ◽  
Author(s):  
Mengwei Sun ◽  
Yuhao Yuan ◽  
Fake Lu ◽  
Anthony J. Di Pasqua
Keyword(s):  
Ovarian Cancer ◽  
Polyethylene Glycol ◽  
Plasmid Dna ◽  
Dna Delivery ◽  
Cationic Liposomes ◽  
Systemic Delivery ◽  
Pegylated Liposome ◽  
Molar Ratios ◽  
Physicochemical Factors ◽  
Insight Into

Cationic liposomes composed of 3-[N-(N’,N’-dimethylaminoethane)-carbamoyl] cholesterol (DC-chol) and dioleoylphosphatidylethanolamine (DOPE) have previously been shown to have applications in gene delivery. Our study aims to explore the effects of inclusion of polyethylene glycol (PEG) and using different molar ratios of DC-chol/DOPE on size, zeta potential, cytotoxicity and DNA delivery of DC-chol/DOPE liposomes. Our results show that PEGylation reduces the cytotoxicity of DC-chol/DOPE liposomes, and, furthermore, PEGylated liposome-DNA lipoplexes are smaller in size and more uniform in size distribution than those that are not PEGylated. Additionally, toxicity against ovarian cancer SKOV-3 cells decreases with the amount of cationic DC-chol present in the formulation; however, decreased delivery of DNA to cellular nuclei is also observed. Transfection with the PEGylated liposomes was successfully demonstrated using plasmid DNA with a known functional outcome. These results offer further insight into physicochemical properties important for cationic liposomes as vehicles for DNA delivery and demonstrate the potential of PEGylated DC-chol/DOPE liposomes as systemic delivery carriers for DNA-mediated ovarian cancer therapy.

scholarly journals Investigation of Plasmid DNA Delivery and Cell Viability Dynamics for Optimal Cell Electrotransfection In Vitro

2020 ◽  
Vol 10 (17) ◽  
pp. 6070
Author(s):  
Sonam Chopra ◽  
Paulius Ruzgys ◽  
Martynas Maciulevičius ◽  
Milda Jakutavičiūtė ◽  
Saulius Šatkauskas
Keyword(s):  
Cell Membrane ◽  
Cell Viability ◽  
High Voltage ◽  
Plasmid Dna ◽  
Transfection Efficiency ◽  
Dna Delivery ◽  
Output Parameter ◽  
Gene Electrotransfer ◽  
Electric Pulses

Electroporation is an effective method for delivering plasmid DNA molecules into cells. The efficiency of gene electrotransfer depends on several factors. To achieve high transfection efficiency while maintaining cell viability is a tedious task in electroporation. Here, we present a combined study in which the dynamics of both evaluation types of transfection efficiency and the cell viability were evaluated in dependence of plasmid concentration as well as at the different number of high voltage (HV) electric pulses. The results of this study reveal a quantitative sigmoidal (R2 > 0.95) dependence of the transfection efficiency and cell viability on the distance between the cell membrane and the nearest plasmid. We propose this distance value as a new, more accurate output parameter that could be used in further optimization studies as a predictor and a measure of electrotransfection efficiency.

scholarly journals Synthesis and Characterization of Chitosan-Saponin Nanoparticle for Application in Plasmid DNA Delivery

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Faruku Bande ◽  
Siti Suri Arshad ◽  
Mohd Hair Bejo ◽  
Shafiu Abdullahi Kamba ◽  
Abdul Rahman Omar
Keyword(s):  
Electron Microscopy ◽  
Delivery System ◽  
Plasmid Dna ◽  
Enzymatic Degradation ◽  
Dna Delivery ◽  
Gravimetric Analysis ◽  
Scanning Calorimetry ◽  
Rapid Decomposition ◽  
Before And After ◽  
Low Cytotoxicity

Nonviral delivery system receives attention over the last decade. Chitosan (CS) is a cationic polymer whereas saponin (SP) is classified as glycoside. In this study, a spherically-shaped CS-SP nanoparticle was synthesized and characterized. The ability of the nanoparticle to protect DNA from enzymatic degradation, its thermostability and cytotoxicity were evaluated. The particle size was found below 100 nm as determined by Zetasizer, transmission electron microscopy (TEM), and field scanning electron microscopy (FSEM) results. The surface charge ranges from 43.7 mV to 38.5 mV before and after encapsulation with DNA plasmid, respectively. In terms of thermostability, Thermal Gravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC) revealed that CS-SP nanoparticle had a melting temperature of 110°C, with rapid decomposition occurring at 120°C. Encapsulation of DNA with the synthesized nanoparticle was evidenced by changes in the FTIR spectra including characteristic peaks at 3267.39 and 1635.58 cm−1, wavenumbers. Additional peak was also observed at 1169.7 cm−1following encapsulation. Electrophoretic mobility showed that CS-SP nanoparticle protected plasmid DNA from enzymatic degradation, while cell viability assays confirmed that the synthesized nanoparticle exhibited low cytotoxicity at different concentrations in avian cells. Taken together these, CS-SP nanoparticle showed potentials for applications as a DNA delivery system.

Peptide-based vectors: recent developments

2014 ◽  
Vol 5 (6) ◽  
pp. 479-488 ◽  
Author(s):  
Carmine Pasquale Cerrato ◽  
Tõnis Lehto ◽  
Ülo Langel
Keyword(s):  
Plasmid Dna ◽  
Cell Types ◽  
Cell Penetrating Peptides ◽  
Nucleic Acid Delivery ◽  
Small Interfering Rnas ◽  
Recent Developments ◽  
Therapeutic Molecules ◽  
Low Cytotoxicity

AbstractPeptides and peptide-cargo complexes have been used for drug delivery and gene therapy. One of the most used delivery vectors are cell-penetrating peptides, due to their ability to be taken up by a variety of cell types and deliver a large variety of cargoes through the cell membrane with low cytotoxicity. In vitro and in vivo studies have shown their possibility and full effectiveness to deliver oligonucleotides, plasmid DNA, small interfering RNAs, antibodies, and drugs. We report in this review some of the latest strategies for peptide-mediated delivery of nucleic acids. It focuses on peptide-based vectors for therapeutic molecules and on nucleic acid delivery. In addition, we discuss recent applications and clinical trials.

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