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High Efficiency Transfection Reagent

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[#GT1211-01] High Efficiency Transfection Reagent


GT1211-01 | High Efficiency Transfection Reagent , 0.5 ml.
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(1) Photoluminescent Cationic Carbon Dots as efficient Non-Viral Delivery of Plasmid SOX9 and Chondrogenesis of Fibroblasts.[TOP]

Pubmed ID :29728593
Publication Date : //
With the increasing demand for higher gene carrier performance, a multifunctional vector could immensely simplify gene delivery for disease treatment; nevertheless, the current non- viral vectors lack self-tracking ability. Here, a type of novel, dual-functional cationic carbon dots (CDs), produced through one-step, microwave-assisted pyrolysis of arginine and glucose, have been utilized as both a self-imaging agent and a non-viral gene vector for chondrogenesis from fibroblasts. The cationic CDs could condense the model gene plasmid SOX9 (pSOX9) to form ultra-small (10-30 nm) nanoparticles which possessed several favorable properties, including high solubility, tunable fluorescence, high yield, low cytotoxicity and outstanding biocompatibility. The MTT assay indicated that CDs/pSOX9 nanoparticles had little cytotoxicity against mouse embryonic fibroblasts (MEFs) compared to Lipofectamine2000 and PEI (25 kDa). Importantly, the CDs/pSOX9 nanoparticles with tunable fluorescence not only enabled the intracellular tracking of the nanoparticles, but also could successfully deliver the pSOX9 into MEFs with significantly high efficiency. Furthermore, the CDs/pSOX9 nanoparticles-mediated transfection of MEFs showed obvious chondrogenic differentiation. Altogether, these findings demonstrated that the CDs prepared in this study could serve as a paradigmatic example of the dual-functional reagent for both self-imaging and effective non-viral gene delivery.

Authors : Cao Xia, Wang Jianping, Deng Wenwen, Chen Jingjing, Wang Yan, Zhou Jie, Du Pan, Xu Wenqian, Wang Qiang, Wang Qilong, Yu Qingtong, Spector Myron, Yu Jiangnan, Xu Ximing,

(2) Cholic acid-modified polyethylenimine: in vitro and in vivo studies.[TOP]

Pubmed ID :29593402
Publication Date : //
Low-molecular-weight polyethylenimine has lower cytotoxicity than high molecular weight polyethylenimine, but it is not an efficient transfection agent because of limitations of DNA delivery into the cytoplasm. Therefore, in the present study, the hydrophobic modification of low-molecular-weight polyethylenimine (PEI 2 kDa [PEI2]) by cholic acid (ChA) was performed to form PEI2-ChA, and in vitro and in vivo studies were performed. Results indicate that the nanoplexes of PEI2-ChA with gWIZ-GFP have greater transfection efficiency (27%) in NT8e cell lines as evaluated by flow cytometry and also observed by fluorescence imaging. The present study concluded that the transferrin-containing nanoplexes of PEI2-ChA conjugates with plasmid p53 warrant clinical trials in humans after exhaustive animal studies for use as a novel gene delivery system.

Authors : Dube Brahmanand, Pandey Abhijeet, Joshi Ganesh, Mulherkar Rita, Sawant Krutika,

(3) Production of Transgenic Mice Through Sperm-Mediated Gene Transfer Using Magnetic Nano-Carriers.[TOP]

Pubmed ID :29490755
Publication Date : //
Current methods of transgenic animal production suffer from low efficiency, cumbersome operation, and high cost. Magnetic nanoparticles (MagNPs) have several characteristics, such as a high carrying efficiency, non-immunogenicity, and strong targeting inducible via magnetic fields, that make them well-suited for use in the generation of transgenic animals. In this study, we used magnetic nano-carriers combined with sperm-mediated gene transfer (SMGT) to generate transgenic mice that harbor the enhanced green fluorescent protein (EGFP) gene. Exogenous plasmid DNA loaded onto Fe3O4 MagNPs were first delivered into mouse sperm cells under a magnetic field. Transfected sperm cells were then incubated with oocytes to complete fertilization, and transgenic mice were successfully generated though embryo transplantation. We demonstrate that this method is exceedingly facile, fast, and cost-effective, with higher transfection efficiency than that of conventional liposome methods.

Authors : Wang Yan, Zhao Xiang, Du Wei, Liu Jinghao, Chen Wenjie, Sun Changjiao, Cui Bo, Zeng Zhanghua, Shen Yue, Gao Fei, Wang Anqi, Liu Guoqiang, Cui Haixin,

(4) Insights into the Influences of Carboxymethyl-β-Cyclodextrin on DNA Formulations Characteristics and Gene Transfection Efficiency.[TOP]

Pubmed ID :29484997
Publication Date : //
Gene therapy is an expanding field and it can treat genetic and acquired diseases.

Authors : Elsana Hassan, Mysina Svetlana, Elkordy Eman Ali, Carr-Wilkinson Jane, Elkordy Amal Ali,

(5) Functionalized Asymmetric Bola-Type Amphiphiles for Efficient Gene and Drug Delivery.[TOP]

Pubmed ID :29462991
Publication Date : //
The studies of bolaamphiphile-based nanoparticles as delivery vectors are still rudimentary and under development. In this study, several asymmetric bolaamphiphiles containing lysine and another moiety with special functions, such as pH-sensitive or cell-targeting property, were designed and synthesized. The potentials of these bolaamphiphile-based nanoparticles as versatile vectors for both nucleic acids and chemical drugs were studied. With the presence of 1,2-dioleoyl--glycero-3-phosphoethanolamine (DOPE), these amphiphiles could be prepared into bolasomes, which showed good DNA binding ability and could condense plasmid DNA into nanoparticles with appropriate size and surface potential. , which has a pH-sensitive histidine on one head, exhibited higher transfection efficiency than the symmetric counterpart and comparable efficiency to commercially available transfection reagent. Mechanism studies confirmed that the bolaplexes formed from might induce the highest cellular uptake and the best endosomal escape ability. On the other hand, these bolaamphiphiles also exhibited good drug loading ability. The self-assembly vesicles could efficiently encapsulate the hydrophobic anti-cancer drug doxorubicin (DOX) in aqueous solution with high drug loading content and encapsulation efficiency. Confocal laser scanning microscopy (CLSM) experiment and cell viability assay exhibited a controlled release of the drug with the assistance of bolasomes. It was shown that such bolaamphiphiles have great potential as nano-vectors for both drug and gene or their co-delivery.

Authors : Huang Zheng, Zhao Dong-Mei, Deng Xuan, Zhang Ji, Zhang Yi-Mei, Yu Xiao-Qi,

(6) Characterization of DNA Condensation by Conformationally Restricted Dipeptides and Gene Delivery.[TOP]

Pubmed ID :29372986
Publication Date : //
A wide variety of non-viral vectors have been developed for gene delivery in past few decades but find limited applications mainly due to lower encapsulation, endosomal entrapment, high toxicity and low transfection efficiency. In this work, we explored plasmid DNA binding ability of several low molecular weight dipeptides containing α,β-dehydrophenylalanine (ΔF) and found that an arginine containing dipeptide, arginine-α,β-dehydrophenylalanine (R-ΔF) condensed pEGFP-N1 plasmid into positively charged spherical nanoparticles of size 250–275 nm. Single molecule techniques showed that R-ΔF interacted with the plasmid DNA in a dose dependent manner which was accompanied by a decrease in diffusion time of the plasmid DNA as well as release of the bound fluorophore. The plasmid DNA in R-ΔF-plasmid complex (R-ΔF-Pl) was stable against DNase action. A pH dependent release of the plasmid DNA from R-ΔF-Pl was observed and the released plasmid DNA retained its natural conformation at endosomal pH, as evidenced from time correlated single photon counting. R-ΔF-Pl was biocompatible and showed ready uptake in HEK 293T cells. Transfection assays using reporter plasmids for green fluorescent protein (GFP), luciferase enzyme and chloramphenicol acetyltransferase (CAT) showed R-ΔF mediated gene delivery both in the presence and absence of serum in the medium. Ease of synthesis, homogenous assembly and biocompatibility balanced with significant expression of gene of interest make R-ΔF a potential vector for development for in vivo application.

Authors : Khatri Anjali, Mishra Aseem, Chauhan Virander Singh,

(7) Proteasome Inhibition Increases the Efficiency of Lentiviral Vector-Mediated Transduction of Trabecular Meshwork.[TOP]

Pubmed ID :29340644
Publication Date : //
To determine if proteasome inhibition using MG132 increased the efficiency of FIV vector-mediated transduction in human trabecular meshwork (TM)-1 cells and monkey organ-cultured anterior segments (MOCAS).

Authors : Aktas Zeynep, Rao Hongyu, Slauson Sarah R, Gabelt B'Ann T, Larsen Inna V, Sheridan Rachael T C, Herrnberger Leonie, Tamm Ernst R, Kaufman Paul L, Brandt Curtis R,

(8) Delivery of expression constructs of secreted frizzled-related protein 4 and its domains by chitosan-dextran sulfate nanoparticles enhances their expression and anti-cancer effects.[TOP]

Pubmed ID :29185158
Publication Date : //
In malignant mesothelioma (MM) cells, secreted frizzled-related protein 4 (SFRP4) expression is downregulated by promoter methylation. In this study, we evaluated the effect of encapsulated chitosan-dextran (CS-DS) nanoparticle formulations of SFRP4 and its cysteine-rich domain (CRD) and netrin-like domain (NLD) as means of SFRP4-GFP protein delivery and their effects in JU77 and ONE58 MM cell lines. CS-DS formulations of SFRP4, CRD, and NLD nanoparticles were prepared by a complex coacervation technique, and particle size ranged from 300 nm for empty particles to 337 nm for particles containing the proteins. Measurement of the zeta potential showed that all preparations were around 25 mV or above, suggesting stable formulation and good affinity for the DNA molecules. The CS-DS nanoparticle formulation maintained high integrity and entrapment efficiency. Gene delivery of SFRP4 and its domains showed enhanced biological effects in both JU77 and ONE58 cell lines when compared to the non-liposomal FUGENE HD transfection reagent. In comparison to the CRD nanoparticles, both the SFRP4 and NLD nanoparticles significantly reduced the viability of MM cells, with the NLD showing the greatest effect. The CS-DS nanoparticle effects were observed at an earlier time point and with lower DNA concentrations. Morphological changes in MM cells were characterized by the formation of membrane-associated vesicles and green fluorescent protein expression specific to SFRP4 and the NLD. The findings from our proof-of-concept study provide a stepping stone for further investigations using in vivo models.

Authors : Perumal Vanathi, Arfuso Frank, Chen Yan, Fox Simon, Dharmarajan Arun M,

(9) Gene delivery ability of polyethylenimine and polyethylene glycol dual-functionalized nanographene oxide in 11 different cell lines.[TOP]

Pubmed ID :29134085
Publication Date : //
We recently developed a polyethylenimine (PEI) and polyethylene glycol (PEG) dual-functionalized reduced graphene oxide (GO) (PEG-nrGO-PEI, RGPP) for high-efficient gene delivery in HepG2 and Hela cell lines. To evaluate the feasibility and applicability of RGPP as a gene delivery carrier, we here assessed the transfection efficiency of RGPP on gene plasmids and siRNA in 11 different cell lines. Commercial polyalkyleneimine cation transfection reagent (TR) was used as comparison. In HepG2 cells, RGPP exhibited much stronger delivery ability for siRNA and large size plasmids than TR. For green fluorescent protein (GFP) plasmid, RGPP showed about 47.1% of transfection efficiency in primary rabbit articular chondrocytes, and about 27% of transfection efficiency in both SH-SY5Y and A549 cell lines. RGPP exhibited about 37.2% of GFP plasmid transfection efficiency in EMT6 cells and about 26.0% of GFP plasmid transfection efficiency in LO2 cells, but induced about 33% of cytotoxicity in both cell lines. In 4T1 and H9C2 cell lines, RGPP had less than 10% of GFP plasmid transfection efficiency. Collectively, RGPP is a potential nano-carrier for high-efficiency gene delivery, and needs to be further optimized for different cell lines.

Authors : Wu Liping, Xie Jinshan, Li Tan, Mai Zihao, Wang Lu, Wang Xiaoping, Chen Tongsheng,

(10) Mussel-inspired polydopamine-polyethylenimine conjugated nanoparticles as efficient gene delivery vectors for mammalian cells.[TOP]

Pubmed ID :29121613
Publication Date : //
Efficient delivery of DNA to cells is the primary concern to address the objective of gene therapy. Many attempts have been made to develop polymeric carriers for gene delivery. To have an efficient carrier, it is vital to understand the properties of the vector for better stability, transfection efficiency and minimal toxicity. Branched polyethylenimine (bPEI) has been considered as the 'gold standard' for gene delivery but suffers a major drawback of exhibiting high cytotoxicity. Here, we have attempted to develop a mussel-derived polymer, polydopamine (PDA), conjugated polyethylenimine nanoparticles in such a way that the toxic nature of bPEI is suppressed by the conversion of free primary amine groups to secondary and tertiary amines. Keeping the amount of PDA fixed, varying amounts of bPEIs of different molecular weights (25, 10 and 1.8kDa) were conjugated via Michael addition and/or Schiff base. A trend in hydrodynamic size of the conjugated nanoparticles was observed in the range from 160 to 300nm and zeta potential from +12-30mV in the projected three series, viz., (i) PDA-bPEI, PDA-bPEI, PDA-bPEI; (ii) PDA-bPEI, PDA-bPEI, PDA-bPEI; and (iii) PDA-bPEI, PDA-bPEI, PDA-bPEI. A visible trend in the DNA condensation ability and buffering capacity was also noticed. Further, cell cytotoxicity assays revealed that pDNA complexes of PDA-bPEI nanoparticles were non-toxic to mammalian cells and these complexes exhibited several folds higher transfection efficiency than the complexes of native bPEIs as demonstrated by fluorescence measurements and flow cytometry. Altogether, the results advocate the promising potential of these conjugated nanoparticles for future in vivo applications.

Authors : Priyam Ayushi, Nagar Prachi, Sharma Ashwani Kumar, Kumar Pradeep,