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Miridian mirna hairpin inhibitor

Manufactured by Horizon Discovery
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Sourced in Germany

MiRIDIAN miRNA hairpin inhibitors are a collection of synthetic oligonucleotides designed to selectively inhibit the activity of specific microRNA (miRNA) molecules. These inhibitors are structured as hairpin-shaped molecules that can bind to and neutralize targeted miRNAs, effectively modulating their regulatory functions within cells.

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Lab products found in correlation

Influx cell sorter, by BD (1 mentions) Mirneasy mini kit, by Qiagen (1 mentions) C fos, by Merck Group (1 mentions) Gata2, by Merck Group (1 mentions) Dotap liposomal transfection reagent, by Merck Group (1 mentions) Stemspan sfem, by STEMCELL (1 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

MiRIDIAN (11 citations) MiRNA inhibitors (4 citations) MiRIDIAN Hairpin Inhibitor (3 citations) MiRNA hairpin inhibitor (2 citations) Miridian miRNA hairpin inhibitors directed against mmu-miR-135b-5p (2 citations) MiRIDIAN miRNA mimics and hairpin inhibitors (2 citations)

5 protocols using miridian mirna hairpin inhibitor

1

Macrophage Differentiation and miRNA Manipulation

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THP-1 cells were differentiated into macrophages at 40°C, harvested and nucleofected with the custom designed 2′-O-methylated miRIDIAN miRNA Hairpin Inhibitor (Dharmacon) directed against hsa-miR-142–5p, hsa-miR-143–3p or non-human miRNA control. These inhibitors do not necessarily direct degradation of miRNAs but would induce loss-of-function as previously published (14 (link)). Cells were co-transfected with pmaxGFP vector and GFP-expressing cells were purified 24 h post nucleofection using fluorescence-activated cell sorting on the InfluxTM Cell Sorter (BD Biosciences). Nucleofection of miR-142 and miR-143 mimics (Dharmacon) and the miRIDIAN microRNA Mimic Negative Control #1 (Dharmacon) was performed using the same protocol. 300nM of miRNA inhibitor or mimic was used in each nucleofection reaction.
Wong J.J., Au A.Y., Gao D., Pinello N., Kwok C.T., Thoeng A., Lau K.A., Gordon J.E., Schmitz U., Feng Y., Nguyen T.V., Middleton R., Bailey C.G., Holst J., Rasko J.E, & Ritchie W. (2016). RBM3 regulates temperature sensitive miR-142–5p and miR-143 (thermomiRs), which target immune genes and control fever. Nucleic Acids Research, 44(6), 2888-2897.
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2

Modulation of miR-224-5p in Cancer Cells

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U2OS osteosarcoma or B16 melanoma cells were seeded at 100,000 cells/well in a 6-well plate. At the following day, cells were either transfected with 300pmol non-targeting control or with miR224-5p. Following 24 hours, the cells were transfected again with a complementary 600pmol specific miR inhibitor (Dharmacon miRIDIAN miRNA Hairpin Inhibitor) plus a negative nonspecific sequence (Dharmacon miRIDIAN miRNA Inhibitor Negative Control). Cells were incubated for 48h hours under normoxic conditions. In the case of hypoxic conditions (1% O2) vs. normoxic conditions - miR224-5p was extracted after 24 hours (B16) or 48 hours (U2OS). In all cases, at the end of incubation microRNA was extracted from the cells by using miRNeasy mini kit (Quiagen). cDNA for miRNA was synthesized from 0.7–1μg RNA by using qScript MicroRNA cDNA Synthesis kit (Quanta).
Silberman A., Goldman O., Assayag O.B., Jacob A., Rabinovich S., Adler L., Sang Lee J., Keshet R., Sarver A., Frug J., Stettner N., Galai S., Persi E., Bahar Halpern K., Zaltsman-Amir Y., Pode-Shakked B., Eilam R., Anikster Y., Nagamani S.C., Ulitsky I., Ruppin E, & Erez A. (2018). Acid-induced downregulation of ASS1 contributes to the maintenance of intracellular pH in cancer. Cancer research, 79(3), 518-533.
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3

Inhibiting miRNA and shRNA Targets

Cited 1 time
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MiRIDIAN miRNA hairpin inhibitors and negative control #1 were obtained from Dharmacon (Lafayette, CO). These miRNA inhibitors bind to miRNAs by complementary sequences and thus block their capacity to silence mRNA target without affecting the level of miRNAs43 (link). Lentiviral particles containing shRNAs against c-Jun (TRCN0000039589, TRCN0000039590), c-Fos (TRCN0000016004, TRCN000016007) and GATA2 (TRCN0000019264, TRCN0000019265) were obtained from Sigma (St Louis, MO).
Zhong L., Huot J, & Simard M.J. (2018). p38 activation induces production of miR-146a and miR-31 to repress E-selectin expression and inhibit transendothelial migration of colon cancer cells. Scientific Reports, 8, 2334.
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4

Ex Vivo Expansion of CD34+ Cells

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CD34+ cells isolated from Cord Blood were seeded at a density of 1.6 × 10^5/well in a 12-well plate and pre-stim for 4 hours in StemSpan SFEM (StemCell Technologies) supplemented with FLT3, SCF, IL-6 and TPO, as indicated. miRIDIAN miRNA hairpin inhibitors (Dharmacon) were transfected using DOTAP Liposomal Transfection Reagent (Sigma-Aldrich). After 24 hours, the media was replaced with fresh media and cells were harvested at 72 hours post-transfection.
Cesana M., Guo M.H., Cacchiarelli D., Wahlster L., Barragan J., Doulatov S., Vo L.T., Salvatori B., Trapnell C., Clement K., Cahan P., Tsanov K.M., Sousa P.M., Tazon-Vega B., Bolondi A., Giorgi F.M., Califano A., Rinn J.L., Meissner A., Hirschhorn J.N, & Daley G.Q. (2018). A CLK3-HMGA2 alternative splicing axis impacts human hematopoietic stem cell molecular identity throughout development. Cell stem cell, 22(4), 575-588.e7.
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5

Inhibition of miRNA in Astrocytes

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Inhibition of rno-miR-31a-5p, -221-3p, -221-5p, -222-3p, -326-3p and -365-3p in cultured astrocytes was performed using miRIDIAN® miRNA hairpin inhibitors (Dharmacon, GE Healthcare Life Sciences, Munich, Germany). Inhibitors were diluted at a final concentration of 40 nmol/l and transfected by using Lipofectamine 2000 reagent (InvitrogenTM, Life Technologies, Darmstadt, Germany) following the protocol of the manufacturer. 6 h after transfection, cell culture media was removed and cells were washed using DMEM (1000 mg/l D-Glucose, GlutaMAXTM) containing 10% FCS following cultivation for another 48 h.
Oenarto J., Karababa A., Castoldi M., Bidmon H.J., Görg B, & Häussinger D. (2016). Ammonia-induced miRNA expression changes in cultured rat astrocytes. Scientific Reports, 6, 18493.
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