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RNotion Transfection Reagent _1 ml

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[#2201500] RNotion Transfection Reagent _1 ml

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(1) MiR-136 triggers apoptosis in human gastric cancer cells by targeting AEG-1 and BCL2.[TOP]

Pubmed ID :30468468
Publication Date : //
Gastric cancer is the second most prevalent cancer across the globe and accounts for about 10% of new cancer cases. It is one among the leading causes of cancer-related deaths around the world. Recently, microRNAs have been identified as important therapeutic targets for the treatment of several cancers owing to their potential to target multiple genes and hinder several biological processes such as proliferation and apoptosis. In the current study, we investigated the potential of miR-136 as therapeutic target for gastric cancer.

Authors : Yu L, Zhou G-Q, Li D-C,

(2) Application of ion pair chromatography coupled with mass spectrometry to assess antisense oligonucleotides concentrations in living cells.[TOP]

Pubmed ID :30462105
Publication Date : //
Antisense oligonucleotides (ASOs) are synthetic bioactive compounds used as therapeutic agents in clinical trials. They act by binding to complementary sequences of the targeted nucleic acids in cells. Assessing the efficiency of ASO delivery to cells or tissues and the stability of these compounds in different biological systems is important. To answer these questions, we developed a new, quick and reliable method to determine the concentrations of different types of ASOs in treated cells. Ultra-high performance liquid chromatography coupled with mass spectrometry was used for the first time for the separation and determination of the studied compounds in total RNA extracts. To develop a method with the highest possible sensitivity, a central composite design was used to comprehensively optimize the MS parameters. Moreover, the effects of the type and concentration of the ion pair reagent on sensitivity were also examined. Finally, a mobile phase containing methanol, hexafluoroisopropanol and N,N-dimethylbutylamine was selected. The optimized method allowed good linearity, accuracy, precision and sensitivity of ASO detection. Next, these compounds were delivered into cells via transfection at a concentration of 25 nM or 125 nM in 1 mL of cell culture medium. After 48 hours, total RNA was isolated from the treated cells and analyzed with the use of the newly developed method. For the cells treated with a higher concentration of ASO composed of phosphorothioate 2'-O-methyl RNA units, the concentration in solution was 0.96 ± 0.06 μM, while in the case of shorter ASO composed of locked nucleic acid units, it was 0.72 ± 0.06 μM in the total RNA extract.

Authors : Studzińska Sylwia, Cywoniuk Piotr, Sobczak Krzysztof,

(3) Establishment of a rapid and footprint-free protocol for differentiation of human embryonic stem cells into pancreatic endocrine cells with synthetic mRNAs encoding transcription factors.[TOP]

Pubmed ID :30359326
Publication Date : //
Transplantation of pancreatic β cells generated in vitro from pluripotent stem cells (hPSCs) such as embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) has been proposed as an alternative therapy for diabetes. Though many differentiation protocols have been developed for this purpose, lentivirus-mediated forced expression of transcription factors (TF)-PDX1 and NKX6.1-has been at the forefront for its relatively fast and straightforward approach. However, considering that such cells will be used for therapeutic purposes in the future, it is desirable to develop a procedure that does not leave any footprint on the genome, as any changes of DNAs could potentially be a source of unintended, concerning effects such as tumorigenicity. In this study, we attempted to establish a novel protocol for rapid and footprint-free hESC differentiation into a pancreatic endocrine lineage by using synthetic mRNAs (synRNAs) encoding PDX1 and NKX6.1. We also tested whether siPOU5F1, which reduces the expression of pluripotency gene POU5F1 (also known as OCT4), can enhance differentiation as reported previously for mesoderm and endoderm lineages.

Authors : Ida Hideomi, Akiyama Tomohiko, Ishiguro Keiichiro, Goparaju Sravan K, Nakatake Yuhki, Chikazawa-Nohtomi Nana, Sato Saeko, Kimura Hiromi, Yokoyama Yukihiro, Nagino Masato, Ko Minoru S H, Ko Shigeru B H,

(4) 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,

(5) 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,

(6) 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,

(7) 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,

(8) 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,

(9) 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,

(10) 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,