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Inherited retinal degeneration (IRD), a group of diseases often causing irreversible blindness, with multiple pathogenesis, still lacks effective treatments currently.Development of effective therapeutics is a primary research goal.Despite rapid advances in gene therapy during the past decades, the most challenging aspect of gene therapy in clinical applications for IRD is to deliver the curative molecules to achieve optimal expression levels in target cells safely.Apart from high gene transfection efficiency, there are still many limitations, such as immunogenicity, biosafety issue, etc.in the application of viral vectors, which drive the development of gene therapy based on non-viral vectors.As one of the hot research topics in non-viral vectors, encouraging progress has been made in DNA nanoparticles for IRD treatment.The polymer/DNA complex nanoparticle is compacted and encapsulated DNA via peptides, lipids, or polysaccharides.Besides, the non-viral delivery system shows cost, preparation, packaging capacity, and safety advantages, providing a promising non-viral platform for safe and effective treatment of IRD, such as retinitis pigmentosa, Stargardt disease, X-linked juvenile retinoschisis, Leber congenital amaurosis, and so on.In this article, advances in transfection efficiency, targeting ability and safety of non-viral gene therapy and its application in IRD were reviewed.
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Poly (β-amino ester)s (PβAEs) contain tertiary amine backbones and biodegradable ester bonds, making them highly biocompatible and pH-responsive. Meanwhile, originated from combinatorial libraries, PβAEs are simple to synthesize, easy to obtain raw materials and can be easily adapted to meet the different performance needs of gene carriers by adjusting the monomer type, monomer ratio and reaction time. Therefore, PβAEs are promising material for non-viral gene carriers. This paper provides a comprehensive overview of the properties and synthesis of PβAEs gene carriers and summarizes the progress of research on the gene delivery of each type of PβAEs.
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Liver fibrosis results from chronic damages together with an accumulation of extracellular matrix, and no specific medical therapy is approved for that until now. Due to liver metabolic capacity for drugs, the fragility of drugs, and the presence of insurmountable physiological obstacles in the way of targeting, the development of efficient drug delivery systems for anti-fibrotics seems vital. We have explored articles with a different perspective on liver fibrosis over the two decades, then collected and summarized the information by providing corresponding and cases. We have discussed the mechanism of hepatic fibrogenesis with different ways of fibrosis induction in animals. Furthermore, the critical chemical and herbal anti-fibrotics, biological molecules such as micro-RNAs, siRNAs, and growth factors, which can affect cell division and differentiation, are mentioned. Likewise, drug and gene delivery and therapeutic systems on and models are summarized in the data tables. This review article enlightens recent advances in emerging drugs and nanocarriers and represents perspectives on targeting strategies employed in liver fibrosis treatment.
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In recent years, non-viral gene vectors have attracted great attention for efficient gene delivery due to the advantages, including low toxicity, low immunogenicity and simple preparation. Polyethylenimine (PEI) is one of the typical non-viral gene carriers that have been widely utilized for gene delivery owing to its superior capabilities in gene compression and buffering capacity. This article discusses the processes of gene delivery and the barriers of PEI-based carrier during the gene delivery, such as low biocompatibility, cytotoxicity, lack of specific targeting and insufficient gene release, etc. Therefore, we summarize the multiple approaches for the modifications of PEI in terms of improved biocompatibility, degradability, specific targeting and buffering capacity. Furthermore, we also review on the recent impressive progresses of smart stimuli-responsive PEI carriers, including endogenous stimuli (pH, reactive oxygen species, glutathione, biomolecular, etc), exogenous stimuli (light, temperature, magnetic field, etc) and dual-responsive strategies, which might provide guidance for the development of more efficient and safer non-viral gene vectors.
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@#Based on the three delivery forms of CRISPR/Cas9 system at the levels of DNA, RNA and protein, this paper mainly approaches the development and new strategies of CRISPR/Cas9 delivery systems, as well as their application in the biomedical field and the clinical treatment of gene-related diseases. By summarizing and elaborating the CRISPR/Cas9 system delivery and gene therapy strategy, new ideas are provided for the discovery of innovative drugs and the development of gene therapy.
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Gene therapy has obvious advantages in the treatment of ocular diseases due to the unique structure of the eye. In recent years, there are more and more therapeutic gene-based drugs for ophthalmic application in clinical trials. Most of the delivery vectors are adeno-associated virus and administered via intraocular injection, which has potential risks. Traditional remedies, such as topical instillationor systemic administration, have limited therapeutic effects on the diseases in the posterior segment of the eye, where the chemical drugs are hard to reach. This makes the research of new strategies for gene drug delivery extremely urgent. For better understanding of the latest hot topics of ocular gene therapy, this article is prepared to introduce application of gene therapy to the typical ocular diseases and the corresponding gene-based medicines. The absorption routes for gene delivery into eyes and existing barriers are summarized. Finally, the gene delivery strategies are highlighted. The clinical application of ocular gene therapy will be boosted by overcoming the absorbing barriers and reducing the potential pitfalls.
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Gene therapy holds a great promise and has been extensively investigated to improve bone formation and regeneration therapies in bone tissue engineering. A variety of osteogenic genes can be delivered by combining different vectors (viral or non-viral), scaffolds and delivery methodologies. Ex vivo & in vivo gene enhanced tissue engineering approaches have led to successful osteogenic differentiation and bone formation. In this article, we review recent advances of gene therapy-based bone tissue engineering discussing strengths and weaknesses of various strategies as well as general overview of gene therapy.
Subject(s)
Bone and Bones , Bone Morphogenetic Proteins , Genetic Therapy , Osteogenesis , Regeneration , Tissue EngineeringABSTRACT
In the recent years ,siRNA has been widely used as effector molecule in the field of gene therapy for cancer . In order to achieve ideal treatment effect ,the key factor is to find safe ,efficient ,stable and controllable gene vectors .The commonly used vectors include viral and non-viral vectors .The non-viral gene vector has superiority of low toxicity ,bare im-munogenicity ,simple to manufacture and high capacity which have been taken as the highlight of the research of carrier of gene drugs .Based on the study of the previous researchers ,this study reviewed the prevalence study of the former carriers in the de-livery system of siRNA from the point of pharmaceutics .
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Objective To study the effects of using 25 kDa linear and branched PEI to transfer plasmid DNA pEGFP -C1 (pDNA ,encoding the enhanced green fluorescent protein reporter gene ) to the basilar membrane of the C57BL/6 mice cochlea in vitro .Methods L -PEI/pDNA and B -PEI/pDNA polyplexes were generated in 0 .1M phosphate buffer solution (PBS) or 5% glucose solution .Polyplexes were characterized by transmission electron mi-croscopy .Agarose gel retardation assay was used to determine the plasmid binding ability of L -PEI and B -PEI . The toxicity was investigated by MTT assay .The transfection was firstly evaluated in 293T cell line ,and then the appropriate amount of PEI and plasmid were applied for cochlear explant transfection of P4 mice pups .Results Un-der the same condition ,B -PEI had better transfection efficiency than L -PEI ,but its toxicity was also higher . When generated in PBS ,the polyplexes had lower toxicity than in glucose solution .L -PEI-pDNA nanoparticles could transfect the spiral limbus fibrocytes ,some spiral ganglion neurons and supporting cells ,but the efficiency was low .Conclusion L -PEI could be used as the non -viral vector for the transfection of the cultured basilar mem-brane of P4 mice pups ,but it should be modified to reach higher efficiency .
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Objective To synthesize miRNA carrier PEG-b-PLL and to testify the stability , encapsulation efficiency and cyto-toxicity of its complexes .Methods H1 NMR was used to determine the degree of polymerization of PLL , 4%agarose gel electrophore-sis was used to determine entrapment of the polyer to the miRNA;then dynamic light scattering ( DLS) was used to measure the hydro-dynamic parameter such as size , polydispersity index ( PDI) and zeta potential of the polyplexes .The entrapment efficiency was deter-mined by ultraviolet-visible spectrophotometer , and finally , the cyto-toxicity of PEG-b-PLL was evaluated by CKK-8 kit with K562 cell lines.Results The characteristics indicated polyplexes prepared by PEG-b-PLL and miRNA fulfill the demand of being the gene carri-er of miRNA because of low cyto-toxicity , high encapsulation efficiency and stability .Conclusion The miRNA carrier PEG-b-PLL had good character and low cyto-toxicity.It showed considerable potential as an efficient miRNA carrier .
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ObjectiveTo investigate the cytotoxicity and gene transfection mediated by NMPCS/DNA nanoparticles.MethodsN-methylene phosphonic chitosan (NMPCS) was synthesized using one-step reaction under homogeneous conditions.The NMPCS/DNA nanoparticles were prepared using complex coacervation method.The cytotoxicity of NMPCS alone and its complexes with plasmid DNA were determined by MTT assay on HeLa cells.The gene transfection mediated by NMPCS/DNA nanoparticles were investigated using pGL3control vector as reporter gene.ResultsThe MTT results suggested that the NMPCS and NMPCS/DNA complexes showed significantly lower cytotoxicity than PEI and PEI/DNA complexes,respectively.The gene transfection mediated by NMPCS/DNA nanoparticles were greatly improved compared with unmodified chitosan.ConclusionNMPCS would demonstrate great potential as a novel,safe,efficient non-viral vector for gene delivery.
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Brain-targeting non-viral gene delivery systems could efficiently mediate gene drugs to cross blood-brain barrier and reach the diseased regions. Many researches have demonstrated that enhanced brain accumulation and gene expression could be achieved via appropriate mechanisms such as specific ligand-receptor binding function. In this review, progress on brain-targeting non-viral gene delivery systems mainly based on receptor- and adsorptive-mediated mechanisms is reviewed.
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Gene vector is defined as a tool which can carry and transfer gene or other nucleic acid material into cells. Its chemical nature can be protein or polypeptide, nucleic acid, lipide, carbohydrate, and other polar molecule or its compound. As an.important part of gene therapy, gene delivery system is also the bottleneck of gene therapy. There are two existent gene vectors defined as viral vectosr and non-viral vectors. Generally, high transfection efficiency can be achieved by the application of viral vectors. But the application of viral vectors has been restricted by its immunogenicity and mutngenic potential as side effects. As a novel gene delivery system, non-viral vectors have the advantages of low toxicity, low immunogenicity, and relative targeting effect. This article provides a review on the latest research progress about various kinds of vectors.
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A 23 amino acid, bifunctional integrin-targeted synthetic oligopeptide was evaluated for ex vivo gene delivery to rabbit bone marrow stromal cells (BMSCs). Synthesis of the peptide (K)16GRGDSPC was performed on a solid-phase batch peptide synthesizer. BMSCs were transfected with plasmid DNA coding for luciferase by (K)16GRGDSPC and the transfection efficiency was assayed. The influences of chloroquine and polyethyleneimine on the transfection efficiency were also examined. The target specificity of (K)16GRGDSPC to mediate exogenous gene into BMSCs was analyzed using cell attachment test and gene delivery inhibition test. The results showed that the transfection efficiency of the oligopeptide vector was lower than that of Lipofectamine. But in the presence of endosomal buffer chloroquine or endosomal disrupting agent polyethyleneimine, the transfection efficiency of the vector was greatly enhanced. In addition, RGD-containing peptides inhibited BMSCs' attachment to the 96-well plates pretreated with fibronectin or vitronectin and significantly decreased the transfection efficiency of the oligopeptide vector. These studies demonstrated that oligopeptide (K)16GRGDSPC was an ideal novel targeted non-viral gene delivery vector, which was easy to be synthesized, high efficient and low cytotoxicity. The vector could effectively deliver exogenous gene into rat BMSCs.
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Objective:To evaluate the feasibility of using polyethyleneimine(PEI) coated magnetic iron oxide nanoparticles(polyMAG-1000) as gene vectors. Methods:The surface characteristies of the nanoparticles were observed with scan electronical microscope.The ability of the nanoparticles to combine and protect DNA were investigated at different PH after the polyMAG-1000 and DNA were combined at different ratio.The nanoparticles were tested as a gene vectors through transfection models in vitro. Results: Under scan electronical microscope, the diameter of the nanoparticles was about 100 nm. The nanoparticles could bind and condense DNA under acidic ,neutral and alkaline pH conditions. The nanoparticles could transfer gene into cell and express green fluorescent protein(GFP).The efficiency of transfection was the highest when the ratio of the nanoparticles and DNA was 1 ∶ 1(v ∶ w).The difference was marked in the transfection efficiency when magnetic field was added or not. Conclusion: The magnetic iron oxide nanoparticles coated with PEI may be potentially used as gene vectors.
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Objective: To investigate the effects of novel targeted non-viral vector in gene therapy of human glioma. Methods: The EGF-R targeting gene delivery system GE7 was constructed. Human Glioma cell line U251 was transfected in vitro with ?－gal as reportor gene and p21 as therapeutic gene using this gene delivery system. By means of the assay of ?-galactosidase staining, Western blotting, in situ end labeling apoptosis cells and DNA ladder, the transferring of exogenous genes and the apoptosis of the tumor cells were examined.Results: It was showed that gene transfer efficiency is over 80%. When transfected with p21 gene, the growth of cells was inhibited significantly, and the apoptosis was detected in the transfected cell by the methods of in situ end labeling and DNA ladder. Conclusion: The GE7 gene delivery system has the ability to transfer exogenous gene to tumor cells and the expression of the therapeutic gene can inhibit the growth of the cells.