scholarly journals The proper strategy to compress and protect plasmid DNA in the Pluronic L64-electropulse system for enhanced intramuscular gene delivery

2019 ◽  
Vol 6 (5) ◽  
pp. 289-298 ◽  
Author(s):  
Yutong He ◽  
Yili Liu ◽  
Zhe Sun ◽  
Fei Han ◽  
James Zhenggui Tang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Delivery ◽  
Natural Compound ◽  
Epigallocatechin Gallate ◽  
Proof Of Concept ◽  
Delivery Method ◽  
Promising Strategy ◽  
Cationic Materials ◽  
Exogenous Gene

Abstract Intramuscular expression of functional proteins is a promising strategy for therapeutic purposes. Previously, we developed an intramuscular gene delivery method by combining Pluronic L64 and optimized electropulse, which is among the most efficient methods to date. However, plasmid DNAs (pDNAs) in this method were not compressed, making them unstable and inefficient in vivo. We considered that a proper compression of pDNAs by an appropriate material should facilitate gene expression in this L64-electropulse system. Here, we reported our finding of such a material, Epigallocatechin gallate (EGCG), a natural compound in green teas, which could compress and protect pDNAs and significantly increase intramuscular gene expression in the L64-electropulse system. Meanwhile, we found that polyethylenimine (PEI) could also slightly improve exogenous gene expression in the optimal procedure. By analysing the characteristic differences between EGCG and PEI, we concluded that negatively charged materials with strong affinity to nucleic acids and/or other properties suitable for gene delivery, such as EGCG, are better alternatives than cationic materials (like PEI) for muscle-based gene delivery. The results revealed that a critical principle for material/pDNA complex benefitting intramuscular gene delivery/expression is to keep the complex negatively charged. This proof-of-concept study displays the breakthrough in compressing pDNAs and provides a principle and strategy to develop more efficient intramuscular gene delivery systems for therapeutic applications.

Gene Expression in Synovial Membrane Cells After Intraarticular Delivery of Plasmid-Linked Superparamagnetic Iron Oxide Particles—A Preliminary Study in Sheep

2006 ◽  
Vol 6 (9) ◽  
pp. 2841-2852 ◽  
Author(s):  
Larry D. Galuppo ◽  
Sarah W. Kamau ◽  
Benedikt Steitz ◽  
Paul O. Hassa ◽  
Monika Hilbe ◽  
...  
Keyword(s):  
Gene Expression ◽  
Iron Oxide ◽  
Gene Delivery ◽  
Synovial Membrane ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Superparamagnetic Iron Oxide ◽  
Oxide Nanoparticles ◽  
Positive Evidence

This study evaluated in vivo gene delivery and subsequent gene expression within cells of the synovium in the presence of static and pulsating magnetic field application following intraarticular injection of superparamagnetic iron oxide nanoparticles linked to plasmids containing reporter genes encoding for fluorescent proteins. Plasmids encoding genes for either green fluorescent protein or red fluorescent protein were bound to superparamagnetic nanoparticles coated with polyethyleneimine. Larger (200–250 nm) and smaller (50 nm) nanoparticles were compared to evaluate the effects of size on transfection efficiency as well as any associated intraarticular reaction. Comparisons between groups were evaluated at 24, 72, and 120 h time periods. Inflammatory response was mild to moderate for all injected particles, but was present in the majority of synovial membrane samples evaluated. Larger particles tended to be associated with more inflammation than smaller ones. Nevertheless, intraarticular application of both experimental and control nanoparticles were well tolerated clinically. Gene expression as determined by observation of either green or red intracellular fluorescence was difficult to assess by both epifluorescent light, and confocal microscopy. An insufficient concentration of nanoparticles in relation to joint volume likely resulted in a limited number of samples with positive evidence of iron staining and with suspected positive evidence of cells expressing fluorescent proteins. Our results indicate that intraarticular administration of functionalized superparamagnetic iron oxide nanoparticles resulted in a mild to moderate synovitis and there was in conclusive evidence of gene expression. Further research is warranted to determine the best and most effective reporter assay for assessment of the in vivo gene delivery into the joints. In addition, the best suited concentration and size of nanoparticles, which will optimize gene delivery and expression, while minimizing intraarticular inflammation, needs to be determined.

scholarly journals Local Gene Delivery System by Bubble Liposomes and Ultrasound Exposure into Joint Synovium

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Yoichi Negishi ◽  
Yuka Tsunoda ◽  
Yoko Endo-Takahashi ◽  
Yusuke Oda ◽  
Ryo Suzuki ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Transfer ◽  
Gene Delivery ◽  
Delivery System ◽  
Imaging System ◽  
Effective Means ◽  
Gene Delivery System ◽  
Highly Efficient ◽  
Efficient Gene

Recently, we have developed novel polyethylene glycol modified liposomes (bubble liposomes; BL) entrapping an ultrasound (US) imaging gas, which can work as a gene delivery tool with US exposure. In this study, we investigated the usefulness of US-mediated gene transfer systems with BL into synoviocytes in vitro and joint synovium in vivo. Highly efficient gene transfer could be achieved in the cultured primary synoviocytes transfected with the combination of BL and US exposure, compared to treatment with plasmid DNA (pDNA) alone, pDNA plus BL, or pDNA plus US. When BL was injected into the knee joints of mice, and US exposure was applied transcutaneously to the injection site, highly efficient gene expression could be observed in the knee joint transfected with the combination of BL and US exposure, compared to treatment with pDNA alone, pDNA plus BL, or pDNA plus US. The localized and prolonged gene expression was also shown by an in vivo luciferase imaging system. Thus, this local gene delivery system into joint synovium using the combination of BL and US exposure may be an effective means for gene therapy in joint disorders.

Human APC gene expression in rodent colonic epithelium in vivo using liposomal gene delivery

1994 ◽  
Vol 3 (11) ◽  
pp. 2005-2010 ◽  
Author(s):  
Carol A. Westbrook ◽  
Steven J. Chmura ◽  
Richard B. Arenas ◽  
Su Young Kim ◽  
Glen Otto
Keyword(s):  
Gene Expression ◽  
Gene Delivery ◽  
Apc Gene ◽  
Colonic Epithelium

scholarly journals The Comparative Utility of Viromer RED and Lipofectamine for Transient Gene Introduction into Glial Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Sudheendra Rao ◽  
Alejo A. Morales ◽  
Damien D. Pearse
Keyword(s):  
Gene Expression ◽  
Gene Delivery ◽  
Glial Cells ◽  
Cell Function ◽  
Viral Vector ◽  
Transient Gene Expression ◽  
Cationic Lipid ◽  
Cell Types ◽  
Nonviral Gene Delivery

The introduction of genes into glial cells for mechanistic studies of cell function and as a therapeutic for gene delivery is an expanding field. Though viral vector based systems do exhibit good delivery efficiency and long-term production of the transgene, the need for transient gene expression, broad and rapid gene setup methodologies, and safety concerns regardingin vivoapplication still incentivize research into the use of nonviral gene delivery methods. In the current study, aviral gene delivery vectors based upon cationic lipid (Lipofectamine 3000) lipoplex or polyethylenimine (Viromer RED) polyplex technologies were examined in cell lines and primary glial cells for their transfection efficiencies, gene expression levels, and toxicity. The transfection efficiencies of polyplex and lipoplex agents were found to be comparable in a limited, yet similar, transfection setting, with or without serum across a number of cell types. However, differential effects on cell-specific transgene expression and reduced viability with cargo loaded polyplex were observed. Overall, our data suggests that polyplex technology could perform comparably to the market dominant lipoplex technology in transfecting various cells lines including glial cells but also stress a need for further refinement of polyplex reagents to minimize their effects on cell viability.

Sustained in Vivo Gene Delivery from Agarose Hydrogel Prolongs Nonviral Gene Expression in Skin

2005 ◽  
Vol 11 (3-4) ◽  
pp. 546-555 ◽  
Author(s):  
Nancy J. Meilander-Lin ◽  
Perrin J. Cheung ◽  
David L. Wilson ◽  
Ravi V. Bellamkonda
Keyword(s):  
Gene Expression ◽  
Gene Delivery ◽  
Agarose Hydrogel ◽  
In Vivo Gene Delivery

The Combination of Drug or Gene Delivery System Responding to Cellular Signals (D-RECS) and Sonoporation System for Effective and Safe Gene Delivery

2009 ◽  
Vol 1237 ◽  
Author(s):  
Akira Tsuchiya ◽  
Takeshi Mori ◽  
Yuki Naritomi ◽  
Jeong-Hun Kang ◽  
Daisuke Asai ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Regulation ◽  
Gene Delivery ◽  
Delivery System ◽  
Transfection Efficiency ◽  
Gene Transfection ◽  
Regulation System ◽  
Gene Delivery System

AbstractWe have developed new gene expression-regulating polymer that can activate transgene expression in response to target intracellular signals. Here, we tried applying sonoporation system to this gene regulation system to enhance the gene expression efficacy. Sonoporation is the method for effective gene transfection in vitro and in vivo. Therefore, the method might enhance the transfection efficiency in our polymer and realize an efficient and safe gene delivery system. Results suggested that the combination of our polymer and sonoporation could improve the gene expression compared to the system using only our polymer that transfers genes into cells via endocytosis. It also kept the ability of the gene regulation responding to cellular signals.

Gene Therapy and Dermatology: More Than Just Skin Deep

1999 ◽  
Vol 3 (5) ◽  
pp. 249-259 ◽  
Author(s):  
Ally-Khan Somani ◽  
Najwa Esmail ◽  
Katherine A. Siminovitch
Keyword(s):  
Gene Expression ◽  
Gene Therapy ◽  
Gene Delivery ◽  
Expression Profiles ◽  
Systemic Disease ◽  
Gene Expression Profiles ◽  
Modulation Of Gene Expression ◽  
Stem Cell Isolation ◽  
Dermatologic Disease

Background: Recent advances in the molecular characterization of dermatologic disease have substantively augmented the understanding of the pathogenetic processes underlying disorders of the skin. This new knowledge coupled with progress in gene delivery technologies has paved the way for introducing cutaneous gene therapy into the dermatologic therapeutic armamentorium. Objective: This review article includes an overview of the current strategies for delivery of gene therapy with an emphasis on the potential role of cutaneous gene delivery in the treatment of skin and systemic diseases. Conclusions: Accessibility for gene delivery, clinical evaluation, and topical modulation of gene expression render the skin a very attractive tissue for therapeutic gene delivery. However, there are several key hurdles to be overcome before cutaneous gene therapy becomes a viable clinical option. These include difficulties in inducing sustained expression of the desired gene in vivo, the challenge of targeting genes to long-lived stem cells, and the difficulty in achieving specific and uniform transfer to different compartments of the skin. However, these problems are not insurmountable and will likely be resolved in conjunction with ongoing advances in delineating gene expression profiles and other molecular properties of the skin, strategies for stem cell isolation, and improved approaches to regulating gene delivery and expression. These advances should create the framework for translating the enormous potential of cutaneous gene therapy into the clinical arena and, thereby, substantively improving the management of both cutaneous and systemic disease.

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