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#29048659   2017/10/19 Save this To Up

MicroRNA-520c-3p negatively regulates EMT by targeting IL-8 to suppress the invasion and migration of breast cancer.

Interleukin-8 (IL-8), which is secreted by cancer cells undergoing epithelial-mesenchymal transition (EMT), can promote EMT in adjacent epithelial-like cells. MicroRNAs (miRNAs/miRs) can affect the expression of target genes via binding to their 3'-untranslated regions (3'-UTRs), which may subsequently affect the biological behaviors of cancer cells. In our previous study, miR-520c-3p was predicted to directly target the 3'-UTR of IL-8. Therefore, the present study was carried out to investigate whether miR-520c-3p can interact with the IL-8 gene and regulate the EMT of breast cancer cells. Web-based prediction algorithms were used to identify miRNAs that potentially target the IL-8 transcript. Luciferase reporter assays were used to confirm the targeting of IL-8 by miR-520c-3p. Reverse transcription-quantitative PCR and western blot analyses were used to examine the levels of IL-8 and EMT-related genes in breast cancer cells. The functional impact of miR-520c-3p on EMT phenotype was evaluated using Transwell and wound-healing assays, and rescue experiments were conducted by overexpressing IL-8 to determine its effect on cell properties. miR-520c-3p was predicted by all three databases, which strongly suggested its interaction with the 3'-UTR of IL-8. The relative Renilla luciferase activity of luciferase reporter construct containing the wild-type 3'-UTR of IL-8 was markedly decreased by miR-520c-3p transfection when compared with scrambled miRNA control transfection (P<0.001). In addition, compared with the scrambled miRNA control transfection, the overexpression of miR-520c-3p significantly reduced the expression of IL-8, and resulted in increased E-cadherin and decreased vimentin and fibronectin levels in MCF-7 and T47D cells (all P<0.001). Introduction of miR-520c-3p inhibited the invasion and migration of MCF-7 and T47D cells (all P<0.001). By contrast, the rescue of IL-8 expression led to the recovery of EMT-related protein expression patterns and cell motility and invasion capabilities. In conclusion, aberrant miR-520c-3p expression may lead to reduced IL-8 expression and promote the mesenchymal phenotype in breast cancer cells, thereby increasing invasive growth.

2633 related Products with: MicroRNA-520c-3p negatively regulates EMT by targeting IL-8 to suppress the invasion and migration of breast cancer.

Breast cancer and normal Advanced breast cancer an Breast cancer and normal Breast cancer tissue arra Middle advanced stage bre Middle advanced stage bre Breast cancer tissue arra Breast cancer tissue arra Breast cancer tissue arra Tissue array of multiple Breast cancer tissue arra Breast cancer tissue arra

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#28986803   2017/10/07 Save this To Up

Knockdown of Rice microRNA166 by Short Tandem Target Mimic (STTM).

Small RNAs, including microRNAs (miRNAs), are abundant in plants and play key roles in controlling plant development and physiology. miRNAs regulate the expression of the target genes involved in key plant processes. Due to functional redundancy among miRNA family members in plants, an ideal approach to silence the expression of all members simultaneously, for their functional characterization, is desirable. Target mimic (TM) was the first approach to achieve this goal. Short tandem target mimic (STTM) is a potent approach complementing TM for silencing miRNAs in plants. STTMs have been successfully used in dicots to block miRNA functions. Here, we describe in detail the protocol for designing STTM construct to block miRNA functions in rice. Such approach can be applied to silence miRNAs in other monocots as well.

2143 related Products with: Knockdown of Rice microRNA166 by Short Tandem Target Mimic (STTM).

NDFIP1 (siRNA target site mFGF 21 (siRNA target sit LacZ (siRNA target site) MOUSE ANTI BOVINE ROTAVIR MOUSE ANTI BORRELIA BURGD Primary antibody FLIP An APC-iFluor™ 700 Tandem APC-iFluor™ 750 Tandem RPE-iFluor™ 750 Tandem Target mRNA Cloning Kit Target mRNA Cloning Kit T Isopeptidase T (short for

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#28986122   2017/10/07 Save this To Up

Mitochondrial ROS-mediated post-transcriptional regulation of α-synuclein through miR-7 and miR-153.

Dysregulation of human alpha-synuclein (α-SYN) is one of the major contributors in the pathogenesis of Parkinson's disease. 1-methyl-4-phenylpyridinium (MPP(+)) is well known neurotoxin which increases α-SYN expression and causes dopaminergic neuronal death. Increasing evidence suggests microRNAs (miRNAs), especially miRNA-7 and miR-153, have important role in the regulation of α-SYN translation and they can prevent MPP(+)-mediated neuronal death. Here, we examined whether MPP(+)-mediated upregulation of α-SYN expression is directly related to miRNA-7 and miR-153. First, we established HEK293/TR cells stably expressing both miR-7 and miR-153. Human α-SYN 3'-UTR containing target sites for both miRNAs was cloned next to a luciferase reporter construct. To control the total levels of reporter mRNA, a tetracycline-inducible system was used. Compared to wild-type HEK293/TR cells, cells overexpressing both miRNAs demonstrated about 75% reduction in luciferase activity. MPP(+) treatment, however, significantly increased luciferase activity of human α-SYN 3'-UTR. Either quenching mitochondrial reactive oxygen species (ROS) or translational inhibition significantly reduced MPP(+)-mediated luciferase activity, suggesting mitochondrial ROS is responsible for MPP(+)-induced α-SYN translation. Together, our results suggest that MPP(+)-mediated increased α-SYN levels are contributed by mitochondrial ROS-mediated de novo protein synthesis which is regulated by miRNA-7 and miR-153.

1018 related Products with: Mitochondrial ROS-mediated post-transcriptional regulation of α-synuclein through miR-7 and miR-153.

∆2-Androstene-1α,17β- Androst-16-en-3-ol C19H30 (5α)-Androst-2-en-17-one (5α)-Androstane-3,11,17- Cell Strainers 70μm Cell DAB (Post) Enhancing Sol DAB (Post) Enhancing Sol Epidermal Growth Factor ( Epidermal Growth Factor ( DNA (cytosine 5) methyltr Active Human Caspase 7100 Androgen Receptor (Phosph

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#28979287   2017/10/05 Save this To Up

An Optimized Transient Dual Luciferase Assay for Quantifying MicroRNA Directed Repression of Targeted Sequences.

Studies investigating the action of small RNAs on computationally predicted target genes require some form of experimental validation. Classical molecular methods of validating microRNA action on target genes are laborious, while approaches that tag predicted target sequences to qualitative reporter genes encounter technical limitations. The aim of this study was to address the challenge of experimentally validating large numbers of computationally predicted microRNA-target transcript interactions using an optimized, quantitative, cost-effective, and scalable approach. The presented method combines transient expression via agroinfiltration of Nicotiana benthamiana leaves with a quantitative dual luciferase reporter system, where firefly luciferase is used to report the microRNA-target sequence interaction and Renilla luciferase is used as an internal standard to normalize expression between replicates. We report the appropriate concentration of N. benthamiana leaf extracts and dilution factor to apply in order to avoid inhibition of firefly LUC activity. Furthermore, the optimal ratio of microRNA precursor expression construct to reporter construct and duration of the incubation period post-agroinfiltration were determined. The optimized dual luciferase assay provides an efficient, repeatable and scalable method to validate and quantify microRNA action on predicted target sequences. The optimized assay was used to validate five predicted targets of rice microRNA miR529b, with as few as six technical replicates. The assay can be extended to assess other small RNA-target sequence interactions, including assessing the functionality of an artificial miRNA or an RNAi construct on a targeted sequence.

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#28978701   2017/10/05 Save this To Up

Processing of Potato Spindle Tuber Viroid (PSTVd) RNAs in Yeast, a Nonconventional Host.

Potato spindle tuber viroid (PSTVd) is a circular, single-stranded, noncoding RNA plant pathogen that is a useful model for the processing of noncoding RNA in eukaryotes. Infective PSTVd circles are replicated via an asymmetric rolling circle mechanism to form linear multimeric RNAs. An endonuclease cleaves these into monomers; a ligase seals these into mature circles. All eukaryotes may have such enzymes for processing noncoding RNA. As a test, we investigated the processing of three PSTVd RNA constructs in the yeast Saccharomyces cerevisiae. Of these, only one form, the construct that adopts a previously described tetraloop-containing conformation (TL), produces circles.TL has 16 nucleotides of the 3' end duplicated at the 5' end and a 3' end produced by self-cleavage of a delta ribozyme. The other two constructs, an exact monomer flanked by ribozymes, and a trihelix-forming RNA with requisite 5' and 3' duplications do not produce circles. The TL circles contain non-native nucleotides resulting from the 3' -end created by the ribozyme and the 5' -end from an endolytic cleavage by yeast at a site distinct from where potato enzymes cut these RNAs. RNAs from all three transcripts are cleaved in places not on path for circle formation, and are likely RNA decay. We propose that these constructs fold into distinct RNA structures that interact differently with host cell RNA metabolism enzymes, resulting in varying susceptibility to degradation versus processing. We conclude that PSTVd RNA is opportunistic and may use different processing pathways in different hosts.IMPORTANCE In higher eukaryotes, the majority of transcribed RNA does not encode proteins. These noncoding RNA are responsible for messenger RNA regulation, control of the expression of regulatory microRNAs, sensing changes in the environment using riboswitches (RNAs that change shape in response to environmental signals), catalysis, and more roles that are still being uncovered. Some of these functions may be remnants from the RNA world, and as such would be part of the evolutionary past of all forms of modern life. Viroids are noncoding RNAs that can cause disease in plants. Since they encode no proteins, they depend on their own RNA and host proteins for replication and pathogenicity. It is likely that viroids hijack critical host RNA pathways for processing the host's own noncoding RNA. These pathways are still unknown. Elucidating these pathways should reveal new biological functions of noncoding RNA.

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Interleukin-34 IL34 (N-t Interleukin-34 IL34 anti HIV1 integrase antibody, Integrin â3 (Phospho Tyr Integrin â3 (Phospho Tyr Integrin â3 (Ab 773) Ant Integrin â3 (Ab 785) Ant Goat Anti-Human ABCE1 RNA Goat Anti-Human RNASEN Dr Polyclonal Antibody Inter Mouse AntiInfluenza B Nuc Mouse AntiInfluenza A Tar

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#28976632   2017/10/04 Save this To Up

miR-CATCH identifies biologically active miRNA regulators of the pro-survival gene XIAP, in Chinese hamster ovary cells.

Genetic engineering of mammalian cells is of interest as a means to boost bio-therapeutic protein yield. X-linked inhibitor of apoptosis (XIAP) overexpression has previously been shown to enhance CHO cell growth and prolong culture longevity while additionally boosting productivity. We confirmed this across a range of recombinant products (SEAP, EPO and IgG). However, stable over-expression of an engineering transgene competes for the cells translational machinery potentially compromising product titre. MicroRNAs are attractive genetic engineering candidates given their non-coding nature and ability to regulate multiple genes simultaneously, thereby relieving the translational burden associated with stable overexpression of a protein-encoding gene. The large number of potential targets of a single miRNA, falsely predicted in-silico, presents difficulties in identifying those that could be useful engineering tools. We explored the identification of direct miRNA regulators of the pro-survival endogenous XIAP gene in CHO-K1 cells by using a miR-CATCH protocol. A biotin-tagged antisense DNA oligonucleotide for XIAP mRNA was designed and used to pull down and capture bound miRNAs. Two miRNAs were chosen out of the 14 miRNAs identified for further validation, miR-124-3p and miR-19b-3p. Transient transfection of mimics for both resulted in the diminished translation of endogenous CHO XIAP protein whereas their inhibition increased XIAP protein levels. A 3'UTR reporter assay confirmed miR-124-3p to be a bona fide regulator of XIAP in CHO-K1 cells. This method demonstrates a useful approach to finding miRNA candidates for CHO cell engineering without competing for the cellular translational machinery.

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DNA (cytosine 5) methyltr anti HSV (II) gB IgG1 (mo anti HCMV IE pp65 IgG1 (m anti HCMV gB IgG1 (monocl Epidermal Growth Factor ( Epidermal Growth Factor ( Human Epstein-Barr Virus Macrophage Colony Stimula Macrophage Colony Stimula Mouse Epstein-Barr Virus Rat TGF-beta-inducible ea Rat TGF-beta-inducible ea

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#28968779   2017/10/02 Save this To Up

A graph regularized non-negative matrix factorization method for identifying microRNA-disease associations.

MicroRNAs (miRNAs) play crucial roles in post-transcriptional regulations and various cellular processes. The identification of disease-related miRNAs provides great insights into the underlying pathogenesis of diseases at a system level. However, most existing computational approaches are biased towards known miRNA-disease associations, which is inappropriate for those new diseases or miRNAs without any known association information.

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#28927049   2017/09/20 Save this To Up

A prognostic model for lung adenocarcinoma patient survival with a focus on four miRNAs.

There is currently no effective biomarker for determining the survival of patients with lung adenocarcinoma. The purpose of the present study was to construct a prognostic survival model using microRNA (miRNA) expression data from patients with lung adenocarcinoma. miRNA data were obtained from The Cancer Genome Atlas, and patients with lung adenocarcinoma were divided into either the training or validation set based on the random allocation principle. The prognostic model focusing on miRNA was constructed, and patients were divided into high-risk or low-risk groups as per the scores, to assess their survival time. The 5-year survival rate from the subgroups within the high- and low-risk groups was assessed. P-values of the prognostic model in the total population, the training set and validation set were 0.0017, 0.01986 and 0.02773, respectively, indicating that the survival time of the lung adenocarcinoma high-risk group was less than that of the low-risk group. Thus, the model had a good assessment effectiveness for the untreated group (P=0.00088) and the Caucasian patient group (P=0.00043). In addition, the model had the best prediction effect for the 5-year survival rate of the Caucasian patient group (AUC=0.629). In conclusion, the prognostic model developed in the present study can be used as an independent prognostic model for patients with lung adenocarcinoma.

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Lung adenocarcinoma and n Lung adenocarcinoma tissu Lung adenocarcinoma (grad Lung adenocarcinoma tissu Lung adenocarcinoma (grad Lung adenocarcinoma tissu Multiple lung carcinoma ( Lung cancer tissue array Lung adenocarcinoma (grad MOUSE ANTI BOVINE ROTAVIR Bone Morphogenetic Protei Growth Differentiation Fa

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#28923913   2017/09/19 Save this To Up

Variations in MicroRNA-25 Expression Influence the Severity of Diabetic Kidney Disease.

Diabetic nephropathy is characterized by persistent albuminuria, progressive decline in GFR, and secondary hypertension. MicroRNAs are dysregulated in diabetic nephropathy, but identification of the specific microRNAs involved remains incomplete. Here, we show that the peripheral blood from patients with diabetes and the kidneys of animals with type 1 or 2 diabetes have low levels of microRNA-25 (miR-25) compared with those of their nondiabetic counterparts. Furthermore, treatment with high glucose decreased the expression of miR-25 in cultured kidney cells. In db/db mice, systemic administration of an miR-25 agomir repressed glomerular fibrosis and reduced high BP. Notably, knockdown of miR-25 in normal mice by systemic administration of an miR-25 antagomir resulted in increased proteinuria, extracellular matrix accumulation, podocyte foot process effacement, and hypertension with renin-angiotensin system activation. However, excessive miR-25 did not cause kidney dysfunction in wild-type mice. RNA sequencing showed the alteration of miR-25 target genes in antagomir-treated mice, including the Ras-related gene CDC42. In vitro, cotransfection with the miR-25 antagomir repressed luciferase activity from a reporter construct containing the CDC42 3' untranslated region. In conclusion, these results reveal a role for miR-25 in diabetic nephropathy and indicate a potential novel therapeutic target for this disease.

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Kidney disease spectrum ( Kidney cancer tissue arra ING1B antisense ING1B sense Interferon γ p19 INK4D AKT1 (dn) Inducible Insulin Insulin promoter factor 1 Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon Anti AGO2 Human, Monoclon

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#28921768   2017/09/18 Save this To Up

MicroRNA-326-3p ameliorates high glucose and ox-LDL-IC- induced fibrotic injury in renal mesangial cells by targeting FcγRIII.

To identify the regulatory relationship between miR-326-3p and FcγRIII, and to explore the involvement of miR-326-3p/FcγRIII/TGF-β/Smad signaling pathway in fibrotic injury, which was induced by the high glucose (HG) and oxidized low density lipoprotein immune complex (ox-LDL-IC) in mouse glomerular mesangial cells (GMCs) METHODS: Dual-luciferase reporter system and real time PCR (RT-PCR) were used to identify FcγRIII as a target gene of miR-326-3p. Lentiviral transduction was utilized to construct different expression of miR-326-3p in GMCs, which were divided into three groups: miR-326-3p mimics group (miR-326-3p group), miR-326-3p inhibitor group (miR-326-3p-inhibit group) and scramble control group (control group). Then, each group was stimulated by normal glucose (NG), HG, ox-LDL-IC and HG+ox-LDL-IC, respectively. RT-PCR and western blot were used to measure the expressions of Col-I, CTGF, α-SMA, TGF-β, Smad2/3 and pSmad2/3.

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Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon GLP 1 ELISA Kit, Rat Gluc Insulin Glucose Phospho-S anti HSV (II) gB IgG1 (mo anti HCMV IE pp65 IgG1 (m anti HCMV gB IgG1 (monocl Human Epstein-Barr Virus Jurkat Cell Extract (Indu Jurkat Cell Extract (Indu

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