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RNotion Transfection Reagent _1 ml

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[#2201500] RNotion Transfection Reagent _1 ml

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(1) Effects of HMGA2 gene downregulation by siRNA on lung carcinoma cell migration in A549 cell lines.[TOP]

Pubmed ID :30317663
Publication Date : //
Although there are multiple treatments for lung cancer, the death rate of this cancer remains high because of metastasis in earlier stages. So a novel treatment for overcoming metastasis is urgently needed. Overexpression of high-mobility group AT-hook 2 (HMGA2), a nonhistone chromosomal protein has been observed in metastatic cancers. So, we suggested that HMGA2 upregulation may play a critical role in treating lung cancer.

Authors : Naghizadeh Sanaz, Mansoori Behzad, Mohammadi Ali, Kafil Hossein Samadi, Mousavi Zohreh, Sakhinia Ebrahim, Baradaran Behzad,

(2) Mechanism of piR-DQ590027/MIR17HG regulating the permeability of glioma conditioned normal BBB.[TOP]

Pubmed ID :30305135
Publication Date : //
The blood-brain barrier (BBB) strongly restricts the entry of anti-glioma drugs into tumor tissues and thus decreases chemotherapy efficacy. Malignant gliomas are highly invasive tumours that use the perivascular space for invasion and co-opt existing vessels as satellite tumor form. Because regulation of the effect of noncoding RNA on BBB function is attracting growing attention, we investigated the effects of noncoding RNA on the permeability of glioma conditioned normal BBB and the mechanism involved using PIWI-associated RNA piR-DQ590027 as a starting point.

Authors : Leng Xue, Ma Jun, Liu Yunhui, Shen Shuyuan, Yu Hai, Zheng Jian, Liu Xiaobai, Liu Libo, Chen Jiajia, Zhao Lini, Ruan Xuelei, Xue Yixue,

(3) High-Throughput DNA Plasmid Transfection Using Acoustic Droplet Ejection Technology.[TOP]

Pubmed ID :30290128
Publication Date : //
The Labcyte Echo acoustic liquid handler allows accurate droplet ejection at high speed from a source well plate to a destination plate. It has already been used in various miniaturized biological assays, such as quantitative PCR (q-PCR), quantitative real-time PCR (q-RT-PCR), protein crystallization, drug screening, cell dispensing, and siRNA transfection. However, no plasmid DNA transfection assay has been published so far using this dispensing technology. In this study, we evaluated the ability of the Echo 550 device to perform plasmid DNA transfection in 384-well plates. Due to the high throughput of this device, we simultaneously optimized the three main parameters of a transfection process: dilution of the transfection reagent, DNA amount, and starting DNA concentration. We defined a four-step protocol whose optimal settings allowed us to transfect HeLa cells with up to 90% efficiency and reach a co-expression of nearly 100% within transfected cells in co-transfection experiments. This fast, reliable, and automated protocol opens new ways to easily and rapidly identify optimal transfection settings for a given cell type. Furthermore, it permits easy software-based transfection control and multiplexing of plasmids distributed on wells of a source plate. This new development could lead to new array applications, such as human ORFeome protein expression or CRISPR-Cas9-based gene function validation in nonpooled screening strategies.

Authors : Colin Béatrice, Deprez Benoit, Couturier Cyril,

(4) Synthesis, characterization and evaluation of transfection efficiency of dexamethasone conjugated poly(propyleneimine) nanocarriers for gene delivery#.[TOP]

Pubmed ID :30270694
Publication Date : //
Polypropylenimine (PPI), a cationic dendrimer with defined structure and positive surface charge, is a potent non-viral vector. Dexamethasone (Dexa) conveys to the nucleus through interaction with its intracellular receptor.

Authors : Malaekeh-Nikouei Bizhan, Rezaee Mehdi, Gholami Leila, Sanjar Mousavi Naghmeh, Kazemi Oskuee Reza,

(5) Transient Gene Expression in Suspension HEK293-EBNA1 Cells.[TOP]

Pubmed ID :30242676
Publication Date : //
Transient gene expression in human embryo kidney 293 (HEK293) cells is an established approach for the rapid production of large amounts of recombinant proteins (r-proteins). Milligram to gram quantities of r-proteins can be typically obtained within less than 10 days following transfection. In this chapter, we describe a simple and robust transfection process of suspension-growing human embryo kidney 293 cells using two commercially available serum-free media and polyethylenimine as the transfection reagent. This chapter provides examples for the production and purification of a his-tagged recombinant protein and two monoclonal antibodies.

Authors : L'Abbé Denis, Bisson Louis, Gervais Christian, Grazzini Eric, Durocher Yves,

(6) HSP90AA1-mediated autophagy promotes drug resistance in osteosarcoma.[TOP]

Pubmed ID :30153855
Publication Date : //
Osteosarcoma is the most common primary bone tumor in children and adolescents. Unfortunately, osteosarcoma treatments often fail due to the development of chemoresistance, of which the underlying molecular mechanisms still remain unclear. In this study, we demonstrated that HSP90AA1 gene is responsible for drug resistance in osteosarcoma through an autophagy-related mechanism.

Authors : Xiao Xin, Wang Wei, Li Yuqian, Yang Di, Li Xiaokang, Shen Chao, Liu Yan, Ke Xianzhu, Guo Shuo, Guo Zheng,

(7) Human wild-type superoxide dismutase 1 gene delivery to rat bone marrow stromal cells: its importance and potential future trends.[TOP]

Pubmed ID :30140407
Publication Date : //
Human superoxide dismutase 1 (SOD1) is the cytosolic form of this enzyme it detoxifies superoxide anions and attenuates their toxicities and concomitant detrimental effects on the cells. It is believed that the amount of these enzymes present in the oxidative stress-induced diseases is crucial for preventing disease progression. Transfection of rat bone marrow stromal cells (BMSCs) by a constructed vector carrying the human wild-type gene, a non-viral gene transfer method, was the main aim of this study.

Authors : Abedi Mohsen, Mesbah-Namin Seyed Alireza, Noori-Zadeh Ali, Tiraihi Taki, Taheri Taher,

(8) Development of an oxidative stress in vitro assay in zebrafish (Danio rerio) cell lines.[TOP]

Pubmed ID :30120374
Publication Date : //
The nuclear factor erythroid 2-related factor 2 (Nrf2) is a key regulator of cellular defense against oxidative stress and correlated with classical toxicological endpoints. In vitro methods using fish cell lines for the assessment of aquatic toxicity are needed for mechanistic studies and as an alternative to in vivo. We describe an in vitro assay to study oxidative stress using zebrafish cell lines. Transfection efficiency of twelve commercially available transfection reagents were tested in the zebrafish cell lines ZFL, ZF4, and Pac2. The most efficient reagent for each cell line was selected for further experiments. Cells were transiently transfected with an Nrf2-responsive luciferase plasmid. The assay was tested using the oxidative stress inducing chemicals tertbutylhydroquinone, hydrogen peroxide, and sulforaphane. Of the transfected cell lines, ZF4 and ZFL showed higher sensitivity. The latter were used to study potential oxidative stress induced by pesticides (diazinon, deltamethrin, atrazine, metazachlor, terbutylazine, diuron). Besides known inducers, Nrf2 activity was also significantly induced by diazinon, deltametrin, diuron, and metazachlor. Activation of Nrf2 by metazachlor is a novel finding. The described assay could be a valuable tool for research in toxicology to study the stress response of both pure chemicals and environmental water samples.

Authors : Lungu-Mitea Sebastian, Oskarsson Agneta, Lundqvist Johan,

(9) Enhanced Intercellular Delivery of cRGD-siRNA Conjugates by an Additional Oligospermine Modification.[TOP]

Pubmed ID :30087937
Publication Date : //
Small interfering RNA (siRNA), consisting a 21-mer duplex molecule, is often modified by conjugation with specific ligands to enhance its capacity for tissue-specific delivery. However, these attempts are hampered by the low permeability of negatively charged RNA molecules to enter the cell membrane. In this study, we designed and synthesized siRNA conjugates modified with cationic oligospermine and cyclic RGD (cRGD) to overcome the low-membrane permeability of siRNA. The siRNA conjugate, which contains 15 spermines and a cRGD peptide, showed sufficient gene-silencing activity at 250 nM final concentration without a transfection reagent. Under these conditions, the cationic oligospermine and cRGD-siRNA conjugate did not show any cytotoxicity.

Authors : Nakamoto Kosuke, Akao Yukihiro, Furuichi Yasuhiro, Ueno Yoshihito,

(10) Transfection reagent Lipofectamine triggers type I interferon signaling activation in macrophages.[TOP]

Pubmed ID :30084169
Publication Date : //
The commercial transfection reagent Lipofectamine has been widely used for cytoplasmic delivery of nucleic acids and for cytosolic engagement with intracellular innate immune sensors to trigger type I interferon (IFN) production. However, the effect of Lipofectamine alone on type I IFN response has not been studied in detail. Here, we show that Lipofectamine induced type I IFN signaling in both RAW 264.7 macrophage-like cells and primary bone marrow-derived macrophages. Type I IFN induction was dependent on interferon regulatory factor (IRF)3 and IRF7 and partially required the toll/interleukin-1 receptor-domain-containing adapter-inducing interferon-β. In contrast, the transfection reagent Xfect did not activate type I IFN signaling. Our study highlights the potential confounding experimental interpretation when using Lipofectamine-based transfection for delivering intracellular ligands and provides important insights into lipid signaling in innate immune responses.

Authors : Guo Xiaomin, Wang Huan, Li Yang, Leng Xiaopeng, Huang Weiwei, Ma Yanbing, Xu Tao, Qi Xiaopeng,