pCMV-SPORT6.1-OPA3 Plasmid

Chickemuel ligand-decorated gold nanoparticles for use in ex vivo modified dendritic cell-based DNA inoculation.

Since mannose receptors (MRs) are expressed on the surfaces of dendritic cells (DCs), the most professional antigen-presenting cells in our body, DNA vaccine vectors containing covalently grafted mannosyl or shikimoyl ligands are used. increasingly on former DNA grafts. live-based transfection. To this end, we have recently shown that ex vivo immunization of mice with cationic amphiphilic liposomes containing 6-aminohexanoic acid in the headgroup region in complex with melanoma antigen-encoded DNA vaccine (MART1) (pCMV-MART1 ) induces long-term immunity. . responses against melanoma (C. Voshavar, et al., J. Med. Chem., 2017, 60, 1605-1610). This finding prompted us to examine, in this investigation, the efficiency of mannose-mimicking chicken ligand (SL)-conjugated gold nanoparticles via a 6-amino hexane thiol spacer (AuNPs-SL) for use in ex vivo-based transfection. gene immunization. Here we report on the design, synthesis, physical and chemical characterization, and bioactivity of AuNPs-SL. Dynamic transmission and light scattering studies by electron microscopy revealed that the hydrodynamic diameters of the AuNPs-SL nanocomparators are in the range of 23 to 44 nm and their surface potentials are in the range of 9 to 28 mV. The MTT assay demonstrated the non-cytotoxic nature of AuNPs-SL and the results of gel electrophoresis assays revealed the strong DNA-binding properties of AuNPs-SL. Importantly, subcutaneous immunization of C57BL/6J mice with DC transfected ex vivo with electrostatic compound of AuNPs-SL and melanoma antigen (MART1) encoding DNA vaccine (p-CMV-MART1) induced durable immunity (100 days). ) against tumor response in immunized mice after subsequent challenge with a lethal dose of melanoma. In particular, mice immunized with ex vivo autologous mbmDC pretransfected with nanoplexes of preformed AuNPs-SL and an unrelated pCMV-SPORT-β-gal plasmid (without the presence of the encoded melanoma antigen) or non-transduced DC, not without durable protection. against subsequent tumor challenge appears. The currently described AuNPs-SL gold nanoparticles (AuNPs-SL) are expected to find future use in DC transfection-based genetic immunization against cancer and other infectious diseases.

A group of cationic lipids based on a benzothiazole head group: synthesis and application for gene delivery.

A series of benzothiazole-based lipids (1-10) containing different benzothiazole derivatives in the main group region were synthesized to determine the relationship between structure and activity for gene delivery. Formulated liposomes were mixed with plasmid DNA encoding green fluorescent protein (α5GFP) or β-galactosidase (pCMV-SPORT-β-gal) and transfected into B16F10 (human melanoma cells), CHO (Chinese hamster ovary), A -549 (human melanoma cells). lung cancer cells) and MCF-7 (human breast cancer cells) types of cancer cells. The efficiencies of Lipids 9 and 10 in particular were found to be comparable and even higher when compared to LipofectAmine-2000. It has been shown to maintain efficient lipid transfection profiles even in the presence of serum. Therefore, the developed benzothiazole headgroup-based lipids have the potential to be used as in vitro and in vivo transfection reagents.

Comparison of the immune responses against foot-and-mouth disease with a DNA vaccine encoding the FMDV/O/IRN/2007 VP1 gene and a conventional inactivated vaccine in an animal model.

Foot and mouth disease virus (FMDV) is highly contagious and is responsible for a large outbreak of disease in cloven-hoofed animals. The objective of this study is to evaluate the plasmid DNA immunization system that expresses the FMDV/O/IRN/2007 VP1 gene and compare it with a conventional inactivated vaccine in an animal model. The VP1 gene was subcloned into the unique KpnI and BamHI cloning sites of the pcDNA3.1+ and pEGFP-N1 vectors to generate the VP gene cassettes (1). BHKT7 cells transfected with the pEGFP-N1-VP1 subcloned vector expressed the GFP-VP1 fusion protein and showed more green dots of fluorescence compared to BHKT7 cells transfected with the pEGFP-N1 vector, which only expresses the GFP protein. Six groups of mice were consecutively immunized with partially cloned PCDNA3.1(+)-VP1 gene cassette as DNA vaccine, PCMV-SPORT-GMCSF vector (as molecular adjuvant) together, conventional vaccine, PBS (as control), and vetion to the DNA vaccine alone, but this response was greater for the combination of conventional vaccines. However, the induction of the brachial immune response in the conventional vaccine group was more protective than in the DNA vaccine, but the proliferation of T cells and the concentration of IFN-γ were higher in the DNA vaccine containing the gene GMCSF. Thus, the group vaccinated with the GMCSF gene DNA vaccine showed a protective neutralizing antibody response and more Th1-type cellular immunity.

A methodology based on the polymerase chain reaction for the detection of gene doping.

The non-therapeutic use of genes to improve sports performance (gene doping) is a new threat to the world of sport. Skeletal muscle is an important target for gene therapy and we asked if we could develop a test system to produce and detect gene doping. To this end, we introduced a plasmid (pCMV-FAK, 3.8 kb, 50 μg) for constitutive expression of the chicken homologue of the muscle growth regulator, focal adhesion kinase (FAK), by electrical gene transfer into M .The single or medial gastrocnemius of the rat. Activation of amplification signals was monitored by assessing muscle fiber cross-section and p70S6K ribosomal kinase. Detectability of the introduced plasmid was monitored by polymerase chain reaction on deoxyribonucleic acid (DNA) from transfected muscle and serum. Muscle transfection with elevated FAK expression of pCMV-FAK 7-fold and 73-fold, respectively, and a mean transverse increase of 52 and 16% in target muscle fibers from soleus muscle and enterococcus muscle after 7 days of gene electrophoresis. The content of p70S6K increased at the same time in the transduced monomuscularis (+110%). Exogenous plasmid sequences could be detected in muscle DNA and cDNA up to 7 days after transfection, but not in serum, except near the site of plasmid deposition, 1 hour after injection and surgery. The results indicate that reliable detection of gene doping in the unethical athlete is not possible unless a change in current tissue sampling practice is implemented, including the collection of muscle biopsies near the gene injection site.


pCMV-SPORT6.1-OPA3 Plasmid

PVT16022 Nova Lifetech 2ug 216 EUR

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PVT46092 Nova Lifetech 2ug 280 EUR

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Hydrodynamic injection of large amounts of plasmid DNA through the hepatic artery results in high-level gene expression in diethylnitrosamine-induced rat hepatocellular carcinomas.

Hydrodynamic injection of naked plasmid DNA (pDNA) via the tail vein is a safe and efficient method of gene transfer to the liver. However, successful gene transfer for hepatocellular carcinoma (HCC) has yet to be demonstrated; Therefore, we investigated the feasibility and efficacy of hydrodynamic injection through the tail vein and hepatic artery in a diethylnitrosamine (DEN)-induced HCC model in mice.
HCC was induced in Sprague-Dawley rats with 100 ppm DEN in drinking water. pCMV-SPORT-beta-galactosidase (beta-gal, 400 μg) (1) was injected via a tail vein at 0.1 mL/g over 30 s or (2) via a hepatic artery at 5 o’clock. 10 mL at 1 mL/s, with or without temporary inferior vena cava (IVC) and portal vein (PV) obstruction. Livers were harvested 24 h after administration and beta-gal expression was assessed by X-gal staining and enzyme activity was measured in tissue homogenates.
Hydrodynamic injection via tail vein achieved gene expression only in non-cancerous tissues (tumor: 0.16 +/- 0.04%, non-tumor: 5.07 +/- 1.66%). Hydrodynamic injection through the hepatic artery was tolerated, but failed to produce efficient gene expression in tumor and non-tumor cells. On the other hand, the simultaneous use of temporary occlusion of IVC/PV with hydrodynamic injection through the hepatic artery significantly increased gene expression in tumor cells, but tumor-selective gene transfer was not achieved with this procedure (tumor: 7, 38 +/- 3.66%, non-tumor: 7.77 +/- 1.06%).
Large-volume hydrodynamic injection of pDNA solution through the hepatic artery with IVC/PV occlusion achieved high-level gene expression in a rat model of HCC. This gene transfer technology may have potential in clinical gene therapy for HCC.

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