BIOCONJUGATE CHEMISTRY
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2024
·
IF 4
Jun Liu,Yanhua Li,Kehui Zhou,Shijia Zhang,Yue Wang,Xiumei Wang,Xiabin Lan,Qixian Chen,Yan Zhao
Electrostatic self-assembly between negatively charged nucleic acids and cationic materials is the basis for the formulation of the delivery systems. Nevertheless, structural disintegration occurs because their colloidal stabilities are frequently insufficient in a hostile biological environment. To overcome the sequential biological barriers encountered during transcellular gene delivery, we attempted to use in situ polymerization onto plasmid DNA (pDNA) with a variety of functional monomers, including N-(3-aminopropyl)methacrylate, (aminopropyl)methacrylamide hydrochloride, 1-vinylimidazole, and 2-methacryloyloxyethylphosphorylcholine and N,N′-bis(acryloyl) cystamine. The covalently linked monomers could polymerize into a network structure on top of pDNA, providing excellent structural stability. Additionally, the significant proton buffering capacity of 1-vinylimidazole is expected to aid in the release of pDNA payloads from acidic and digestive endolysosomes. In addition, the redox-mediated cleavage of the disulfide bond in N,N′-bis(acryloyl)cystamine allows for the selective cleavage of the covalently linked network in the cytosolic microenvironment. This is due to the high intracellular level of glutathione, which promotes the liberation of pDNA payloads in the cell interiors. The proposed polymerization strategies resulted in well-defined nanoscale pDNA delivery systems. Excellent colloidal stabilities were observed, even when incubated in the presence of high concentrations of heparin (10 mg/mL). In contrast, the release of pDNA was confirmed upon incubation in the presence of glutathione, mimicking the intracellular microenvironment. Cell transfection experiments verified their efficient cellular uptake and gene expression activities in the hard-transfected MCF-7 cells. Hence, the polymerization strategy used in the fabrication of covalently linked nonviral gene delivery systems shows promise in creating high-performance gene delivery systems with diverse functions. This could open new avenues in cellular microenvironment engineering.
研究领域:
生物材料学
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基因治疗
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药物递送系统
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHYSICAL CHEMISTRY B
·
2014
·
IF 3.377
Long-Hai Wang,Sheng-Gang Ding,Jun-Jie Yan,Ye-Zi You
Encapsulation of negatively charged plasmid DNA into a small-sized nanocapsule without using any condensing agent is very challenging up to now. Here we report a versatile method for encapsulating large-sized plasmid DNAs into small-sized bioreducible nanocapsules in which shearing force and surfactant can fold large-sized plasmid DNAs into small-sized emulsion droplets containing bioreducible branched polymers. Subsequently, temperature triggers the bioreducible branched polymers to aggregate and cross-link at the water/oil interface of the emulsion nanodroplet, forming a bioreducible shell around the nanodroplet. Thus, a small-sized nanocapsule (∼110 nm) containing large-sized plasmid DNA (∼1900 nm long) forms by removal of the surfactant.
研究领域:
生物材料学
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基因治疗
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药剂学
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DOI
高影响因子
Journal of Nanomaterials
·
2017
·
IF 1.871
Juan Zhang,Huanhuan Weng,Xiangwan Miao,Quanming Li,Siqi Wang,Huifen Xie,Tao Liu,Minqiang Xie
A novel folate (FA) receptor-targeted superparamagnetic Fe3O4 nanoparticles (SPIONs) codelivering cisplatin (CDDP) and tissue factor pathway inhibitor-2 (TFPI-2) plasmid DNA (pDNA) was constructed. The core shell nanocomposites ([email protected]) were composed of superparamagnetic Fe3O4 core that binds CDDP and TFPI-2 shell that combines with folate-polyethylene glycol-polyethyleneimine (FA-PEG-PEI) via electrostatic interaction. The shell containing FA-PEG-PEI and TFPI-2 plasmid was synthesized through amidation reaction and electrostatic adsorption and the core containing SPION-CDDP was modified by aldehyde sodium alginate. Proton nuclear magnetic resonance and Fourier transform infrared spectra on FA-PEG-PEI polymers showed characteristic peaks of various metabolites in corresponding frequency. Transmission electron microscopy image of [email protected] nanoparticles demonstrated a near-monodisperse spherical morphology, while dynamic light scattering studies indicated an intensity-average diameter of 149.5 nm. Zeta potential was 14.89 ± 1.83 mv and the final concentration of loaded CDDP was 100 ug/ml. Gel electrophoresis data showed that the nanocomposite would protect TFPI-2 pDNA from being digested by DNases. Compared with CNE-2 cells, the good targetability and better gene transfection efficiency (57.9%) were detected by Prussian blue iron stain and fluorescence analysis in HNE-1 cells in vitro. The results suggested the potential application of [email protected] as a multifunctional anticancer nanomedicine on targeting therapy for FR positive NPC.
研究领域:
纳米技术
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癌症治疗
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药学
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生物医学工程
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产品关联
DOI
高影响因子
STARCH-STARKE
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2015
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IF 1.677
Xuan Liang,Shuyun Wu,Xianyue Ren,Daping Quan,Li-Ming Zhang,Yubing Deng,Liqun Yang
A series of hyperbranched glycogen derivatives conjugated with different contents of cationic 3-(dimethylamino)-1-propylamine residues, named DMAPA-Glyp derivatives, were synthesized at room temperature. Resonance light scattering (RLS) spectroscopy was used to investigate the complexation behavior and efficacy of the DMAPA-Glyp derivatives and plasmid DNA (pDNA). Results indicated that the DMAPA-Glyp derivatives and pDNA molecules began to aggregate and form complexes at a weight ratio of 2. Following further increases in the DMAPA-Glyp derivatives (w/w >2), pDNA was condensed to form compact complexes. RLS spectroscopy is thus a useful tool for studying the complexation behavior and efficacy of cationic polymers with nucleic acid molecules. Circular dichroism analysis further revealed that the conformation of pDNA transformed from B-form to C-form and A-form during the complexation process.
研究领域:
生物化学
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分子生物学
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产品关联
DOI
高影响因子
CHEMICAL ENGINEERING JOURNAL
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2020
·
IF 10.652
Renkuan Liao,Jiaojiao Zhang,Taotao Li,Dan Luo,Dayong Yang
Pollutants being discharged into water body from unknown sources are one of the major threats to environmental sustainability and human health. Herein a plasmid DNA (pDNA)-based tracing system for multiplexed detection of potential hydrological processes of pollutants is developed. pDNA is incorporated with magnetic nanoparticles (MNPs) and then the pDNA-MNPs nanocomposites are encapsulated inside polylactic acid (PLA) microspheres as pDNA tracers. pDNA, produced in large quantities by engineering bacteria ( E. coli ), served as the “marker” which can be arbitrarily designed and specifically identified through well-established molecular biology tools including enzyme. MNPs are employed to facilitate the collection and separation of tracers in diluted water body. PLA microspheres provide effective protections for pDNA against unpredictable influences in environment such as high temperature, UV radiation, and deoxyribonuclease degradation. In addition, pDNA tracers containing MNPs are buoyant and concentrated at the surface of the water, facilitating tracer collection with an external magnetic device. pDNA extracted from tracers is specifically detected via quantitative Polymerase Chain Reaction (qPCR) with extreme sensitivity. Multiplexed detection is also achieved both in a laboratory test and in a simulated-stream test. Moreover, the preliminary application strategies of pDNA tracer system in different scales of a real-world river have been envisioned. The advantages of pDNA-based tracing system include: massive and cost-effective production of pDNA, high stability in harsh environment, environmental friendly, ultrasensitive, and allowing for multiplexed detection.
研究领域:
环境科学
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分子生物学
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纳米技术
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产品关联
DOI
高影响因子
Cellular and Molecular Bioengineering
·
2019
·Vol. 13
·期 1
·99-112
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IF 2.477
Wang Yue,Li Xiaoli,Liu Lanlan,Liu Bingruo,Wang Feng,Chen Changsheng
Introduction Ultrasound-targeted microbubble destruction (UTMD) has been shown a promising approach for target-specific gene delivery and treatment of many diseases in the past decade. To improve the therapeutic potential of UTMD, the gene carrier of microbubbles should possess adequate DNA condensation capability and (or) specific cell or tissue selectivity. The tissue-targeted and ultrasound-targeted cationic microbubbles were developed to meet gene therapy. Methods A tissue-targeted stearic acid-inserted cationic microbubbles (SCMBs) were prepared for ultrasound-targeted gene delivery. Branched PEI was modified with stearic acid and further mixed with 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and biot-1,2-distearoyl-sn-glycero-3-phosphoethanolamine- N -[methoxy (polyethylene glycol)-2000] (ammonium salt) (Biot-DSPE-PEG2000), intercellular adhesion molecule-1 (ICAM-1) antibody and plasmid DNA to prepare cationic microbubbles through ultrasonic hydration. The ICAM-1 antibody and plasmid DNA were expected to assemble to the surface of SCMBs via biotin-avidin interaction and electrostatic interaction, respectively. Results It was found that the SCMBs had higher zeta potential compared with neutral microbubbles (NMBs) and cationic microbubbles (CMBs). In contrast, DNA incorporated SCMBs4 showed negative potential, exhibiting good DNA-binding capacity. Confocal images showed that the HeLa cells were attached around by the SCMBs4 from the view of green fluorescence of fluorescein isothiocyanate-loaded IgG which conjugated to ICAM-1 antibody on their surface. After ultrasound treatment, HeLa cells treated with SCMBs exhibited slightly stronger red fluorescence under confocal laser scanning microscope, indicating a synergistic promotion for transfection efficiency. Conclusions This tissue- and ultrasound-targeted cationic microbubble demonstrated here showed a promising strategy for improving gene therapy in the future.
研究领域:
生物医学工程
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基因治疗
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药学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
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2024
·Vol. 22
·期 1
·1-15
·
IF 10.6
Wang Xiaobo,Cai Chengyuan,Lv Weiqi,Chen Kechen,Li Jiaxin,Liao Kaitong,Zhang Yanqun,Huang Hongxin,Lin Ying,Rong Zhili,Duan Xiaopin
CRISPR-based gene therapy offers precise targeting and specific editing of disease-related gene sequences, potentially yielding long-lasting treatment effects. However, efficient delivery remains a significant challenge for its widespread application. In this study, we design a novel short peptide-conjugated bioreducible polymer named TSPscp as a safe and effective delivery vector for the CRISPR system. Our results show that TSPscp markedly boosts transcriptional activation and genome editing activities of multiple CRISPR systems as confirmed by decomposition-seq and Deep-seq, which is resulted from its capability in facilitating delivery of plasmid DNA by promoting cellular uptake and lysosomal escape. Additionally, TSPscp further enhances genome editing of CRISPR by delivery of minicircle DNA, a condensed form of regular plasmid DNA. More importantly, TSPscp significantly improves delivery and genome editing of CRISPR system in vivo. In summary, our study highlights TSPscp as a promising delivery tool for CRISPR applications in vivo.Graphical Abstract
研究领域:
基因治疗
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分子生物学
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生物技术
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产品关联
DOI
PubMed
高影响因子
MACROMOLECULAR BIOSCIENCE
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2017
·Vol. 17
·期 8
·1700114
·
IF 3.238
Ting Wu,Longhai Wang,Shenggang Ding,Yezi You
A novel PEGylation polypeptide, poly(ethylene glycol)- b -poly( l -lysine)- b -poly( l -cysteine) (PEG-PLL-PCys) triblock copolymer is synthesized via the sequential ring-opening polymerization of amino acid N -carboxyanhydrides initiated by methoxypolyethylene glycol amine (mPEG-NH 2 , M w is 2 kDa). Subsequently, the obtained polypeptide is partially conjugated with fluorocarbon chains via disulfide exchange reaction. PLL segment can condense plasmid DNA through an electrostatic force to form a complex core, PEG segment surrounding the complex like a corona can prevent the complex from precipitation and reduce the adsorption of serum, while PCys segment with fluorocarbon can enhance the cellular uptake and the stability of the formed polyplex micelles in physiological conditions. Experiment results exhibit that the fluorinated polypeptides have low cytotoxicity and good gene transfection efficiency even in the presence of 50% fetal bovine serum.
研究领域:
高分子化学
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药学
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生物医学工程
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DOI
PubMed
高影响因子
BIOORGANIC & MEDICINAL CHEMISTRY
·
2013
·
IF 2.903
Ruilong Sheng,Ting Luo,Hui Li,Jingjing Sun,Zhao Wang,Amin Cao
In this work , we have successfully prepared a series of new sterol-based cationic lipids ( 1 – 4 ) via an efficient ‘Click’ chemistry approach. The pDNA binding affinity of these lipids was examined by EB displacement and agarose-gel retardant assay. The average particle sizes and surface charges of the sterol-based cationic lipids/pDNA lipoplexes were analyzed by dynamic laser light scattering instrument (DLS), and the morphologies of the lipoplexes were observed by atomic force microscopy (AFM). The cytotoxicity of the lipids were examined by MTT and LDH assay, and the gene transfection efficiencies of these lipid carriers were investigated by luciferase gene transfection assay in various cell lines. In addition, the intracellular uptake and trafficking/localization behavior of the Cy3-DNA loaded lipoplexes were preliminarily studied by fluorescence microscopy . The results demonstrated that the pDNA loading capacity, lipoplex particle size, zeta potential and morphology of the sterol lipids/pDNA lipoplexes depended largely on the molecular structure factors including sterol-skeletons and headgroups. Furthermore, the sterol-based lipids showed quite different cytotoxicity and gene transfection efficacy in A549 and HeLa cells. Interestingly, it was found that the cholesterol-bearing lipids 1 and 2 showed 7–10 4 times higher transfection capability than their lithocholate-bearing counterparts 3 and 4 in A549 and HeLa cell lines , suggested that the gene transfection capacity strongly relied on the structure of sterol skeletons . Moreover, the study on the structure–activity relationships of these sterol-based cationic lipid gene carriers provided a possible approach for developing low cytotoxic and high efficient lipid gene carriers by selecting suitable sterol hydrophobes and cationic headgroups.
研究领域:
分子生物学
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基因治疗
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药剂学
✓ 已验证
DOI
PubMed
高影响因子
Journal of Radiation Research and Applied Sciences
·
2020
·
IF 1.804
Xue-Wen Liu,Ning-Yi Liu,Yuan-Qing Deng,Shan Wang,Ting Liu,Yu-Cai Tang,Yuan-Dao Chen,Ji-Lin Lu
ABSTRACT In recent years, ruthenium (II) complexes have attracted attention due to high DNA topoisomerase I inhibitory activities and good anticancer activities. Herein, an asymmetric ligand 2-(5-(1, 10-phenanthroline))-1 H-4,5-diphenyl-imidazole (pdpi) and its ruthenium complexes were synthesized. DNA-binding mode of two complexes was determined as intercalation by using UV–vis spectra, emission spectra, viscosity, and molecular docking experiments. DNA photocleavage experimental results show that two ruthenium complexes effectively cleave plasmid DNA by producing singlet oxygen. Furthermore, they both displayed good topo I inhibition activities. The cytotoxic activities results show that complex 2 displays better antitumor activities against Eca-109 cells and A549 cells (IC 50 = 25.97 ± 3.33 μM and 28.83 ± 2.49 μM, respectively) compared to complex 1 .
研究领域:
化学
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药理学
·
癌症研究
✓ 已验证
DOI
高影响因子
Journal of Materials Chemistry B
·
2016
·Vol. 4
·期 7
·1284-1295
·
IF 4.872
Min Tang,Haiqing Dong,Yongyong Li,Tianbin Ren
Dendrimer catiomers like dendrigraft poly-L-lysine (DGL) have been very popular vectors for gene delivery recently; however, they generally suffer from serious cytotoxicity for high density of positive charge. PEGylated DGL engineered using the PEG cleavable mechanism (DGL(R)-SS-mPEG) was first developed as a non-viral gene vector for cancer intervention. Cleavable PEGylation of the DGL catiomer in tumor relevant glutathione (GSH) conditions enables us to dramatically decrease the cytotoxicity as well as to promote the intracellular release and expression of the genetic payload. Like DGL, DGL(R)-SS-mPEG is capable of efficiently complexing with plasmid DNA (pDNA) to afford homogeneous compact nano-complexes. Those gene carrying nanostructures could be stably dispersed in the regular serum medium without GSH, but with fast PEG dis-assembly if subject to 10 mM GSH. Compared with the non-cleavable counterpart, PEG-cleavable dendrigraft poly-L-lysine exhibited significantly higher enhanced green fluorescence protein (EGFP) expression against 293T cells. By using small interfering RNA (siRNA-VEGF) as the therapeutic gene payload, the complex nanoparticles demonstrated the pronounced inhibition effect on cell growth in vitro and tumor growth in vivo. The promising results revealed a universal strategy to balance disadvantages and advantages of dendrimer catiomers for future non-viral gene delivery vector.
研究领域:
药学
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生物技术
·
基因治疗
·
癌症研究
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产品关联
DOI
PubMed
高影响因子
Journal of Materials Chemistry B
·
2014
·Vol. 3
·期 8
·1507-1517
·
IF 0
Xiaojun Cai,Yongyong Li,Dong Yue,Qiangying Yi,Shuo Li,Donglu Shi,Zhongwu Gu
Gene carriers made from polylysine are of interest in relation to gene therapy but suffer from the lack of transfection efficiency due to limited stability and endosomal escape ability. To address this problem, we designed and developed Schiff-base linked imidazole modified polylysine with a reversible-PEGylation catiomer (SL-ImPEG-SS-PLL) for high efficiency gene delivery. The reversible PEGylation was introduced for in vivo circulation, as well as selective PEG detachment to augment the cellular internalization, while introducing Schiff-base linked imidazole residues into polylysine was expected to accelerate the endosomal escape of the DNA payload, as well as facilitate intracellular DNA unpacking and release, thus significantly enhancing gene delivery efficiency. The size alteration of the SL-I15mPEG-SS-PLL/pDNA polyplexes in the presence of 10 mM GSH suggests stimulus-induced PEG detachment under tumor relevant reduction conditions. Acid–base titration assays indicate that imidazole residues confer on polylysine remarkable buffering ability. The agarose gel retardation assay suggests that the Schiff-base linkages provide an increased DNA binding ability to protect DNA against nucleases and timely intracellular DNA unpacking to permit DNA dissociation from polylysine and access to transcriptional machinery. Biological efficacy assessment of this multifunctional carrier, using pEGFP and pGL-3 as reporter genes, indicates comparable to or even higher transfection efficiencies than gold standard PEI and their transfection efficiencies are slightly affected by serum. More importantly, in vivo transfection of pEGFP reveals that GFP expression was found not only in some of the important organs, such as the liver, spleen, kidneys and lungs, but also in the transplanted carcinoma. These experimental results suggest that the reversible PEGylation and Schiff-base linked imidazole modification make SL-ImPEG-SS-PLL a great potential candidate for an effective and biocompatible gene delivery system.
研究领域:
生物技术
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基因治疗
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药学科学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
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