Anti mir 146a

Manufactured by Thermo Fisher Scientific

Sourced in Germany

Anti-miR-146a is a laboratory reagent designed to inhibit the function of miR-146a, a microRNA involved in various cellular processes. It can be used in research applications to study the role of miR-146a in biological systems.

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Lab products found in correlation

Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Mir 146a inhibitor, by GenePharma (1 mentions) Mir 146a mimic, by GenePharma (1 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions) Anti mir 155, by Thermo Fisher Scientific (1 mentions) Pmaxgfp, by Lonza (1 mentions) 4d nucleofector system, by Lonza (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

MiR-146a Anti-miRs

4 protocols using anti mir 146a

Modulation of miR-146a and CNTFR in Leukemia Cells

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Jurkat and HL-60 cells were added to 6-well plates and incubated at 37℃ for 24 h. When cells reached 80% confluence in the plate well, anti-miR-146a (antisense miR-146a oligonucleotide, Thermo), anti-miR-146a negative control (NC, Thermo), CNTFR-siRNA (QIAGEN, Duesseldorf, Germany), CNTFR-siRNA negative control (QIAGEN), miR-146a mimics (GenePharma, Shanghai, China), miR-146a mimics negative control (GenePharma), and miR-146a inhibitor (GenePharma) were cotransfected into Jurkat and HL-60 cells using Lipofectamine® 2000 Reagent (Invitrogen). The transfected Jurkat and HL-60 cells were randomly assigned to eight groups: Mock group (no treatment), mimics-NC group (transfected with miR-146a mimics negative control), miR-146a mimics group (transfected with miR-146a mimics), miR-146a inhibitor group (transfected with miR-146a inhibitor), anti-miR-NC+siNC group (transfected with anti-miR-146a NC and CNTFR-siRNA NC), anti-miR-146a+siNC group (transfected with anti-miR-146a and CNTFR-siRNA NC), anti-miR-NC+siCNTFR group (transfected with anti-miR-146a NC and CNTFR-siRNA), and anti-miR-146a+siCNTFR group (transfected with anti-miR-146a and CNTFR-siRNA). Finally, all cells were cultured in 37℃ incubator for 48 h.

Wang L., Zhang H, & Lei D. (2019). microRNA-146a Promotes Growth of Acute Leukemia Cells by Downregulating Ciliary Neurotrophic Factor Receptor and Activating JAK2/STAT3 Signaling. Yonsei Medical Journal, 60(10), 924-934.

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miR146a Activity Inhibition Technique

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miR146a activity was inhibited by using simultaneously two anti-miR146a, targeting the 3p and the 5p (Thermo Fisher Scientific). Such anti-miRNAs were transfected into the cell lines through Lipofectamine 3000 (Thermo Fisher Scientific) according to the manufacturer’s instruction. According to our optimized conditions, a final concentration of 30 nM was enough to inhibit miR146a activity for at least 72 h. Higher concentrations (a maximum of 100 nM was tested) did not exert better or long-lasting effects.

Biagioni A., Chillà A., Del Rosso M., Fibbi G., Scavone F., Andreucci E., Peppicelli S., Bianchini F., Calorini L., Li Santi A., Ragno P., Margheri F, & Laurenzana A. (2021). CRISPR/Cas9 uPAR Gene Knockout Results in Tumor Growth Inhibition, EGFR Downregulation and Induction of Stemness Markers in Melanoma and Colon Carcinoma Cell Lines. Frontiers in Oncology, 11, 663225.

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Electroporation-Mediated Anti-miR Transfection

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Anti-miR transfections were performed by electroporation (Nucleofector 4D System; Lonza Cologne AG, Cologne, Germany). MUM2B and OCM1 cells (5 × 10⁶) were suspended in transfection medium consisting of 100 ul DMEM with 5 mM KCl, 15 mM MgCl₂, 15 mM HEPES, and 150 mM Na₂HPO₄/NaH₂PO₄ (pH 7.2), and anti-miR-155 and anti-miR-146a (Ambion, Austin, TX) were added (1 ul of 5 uM solution). Transfection was accomplished with the FF-120 pulse. Cells electroporated without anti-miRs were used as controls. Cells were then incubated in DMEM with 10% heat-inactivated fetal calf serum cells for 1 hour prior to testing. Transfection efficiency was monitored using a co-transfected GFP-expressing plasmid (Pmax GFP, Lonza).

Joshi P., Kooski M., Aldrich W., Varghai D., Zborowski M., Singh A.D, & Triozzi P.L. (2016). Expression of natural killer cell regulatory microRNA by uveal melanoma cancer stem cells. Clinical & experimental metastasis, 33(8), 829-838.

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Silencing miR-146a in Synovial Fibroblasts

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Synovial fibroblasts were transfected with 100 picomoles of anti-miR-146a (Ambion) and Cy3-labeled control anti-miR (Ambion) with Lipofectamine 2000 as per the manufacturer’s protocols. Efficiency of transfection was monitored by visualizing the fluorescence of Cy3-labeled control anti-miR. Cells were harvested for RNA isolation and protein lysate preparation 48 hours post transfection. Knockdown of miR-146a in anti-miR transfected cells were confirmed by quantitative real time PCR using TaqMan assay specific to miR-146a. Protein expression levels of miR-146a targets were analysed in the transfected cells by Western blotting with the corresponding antibodies.

Selvamani S.P., Mishra R, & Singh S.K. (2014). Chikungunya Virus Exploits miR-146a to Regulate NF-κB Pathway in Human Synovial Fibroblasts. PLoS ONE, 9(8), e103624.

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