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Control mirna mimic

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The Control miRNA mimic is a synthetic, non-targeting small RNA molecule designed to serve as a negative control in miRNA expression experiments. Its core function is to provide a reference point for evaluating the effects of experimental miRNA mimics on gene expression and cellular processes.

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Lab products found in correlation

J 028584 12 0020, by Horizon Discovery (2 mentions) Let 7a mirna mimic, by Thermo Fisher Scientific (2 mentions) Let 7a inhibitor, by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Dual luciferase reporter assay system, by Promega (2 mentions) Pgl3 control vector, by Promega (2 mentions) Amaxa nucleofector system, by Lonza (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

MiRNA mimics (152 citations) Control mimic (31 citations) Control miRNA (26 citations) MirVana miRNA Mimic miR-1 Positive Control (4 citations) MirVana mimic miRNA control #1 (3 citations)

6 protocols using control mirna mimic

1

Reverse Transfection and Functional Assays

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BE(2)C and Kelly cells were reverse transfected using Lipofectamine 2000© (Life Technologies) into 6 well plates using the appropriate siRNA or miRNA mimics (described below). Cells were harvested at time-points described for analysis by western or qPCR. Growth assays were performed similarly, but in 96 well plates followed by time-point specific BrdU growth assay. Global let-7 target analysis: BE(2)C:MYCN-ORF cells co-transfected with control or MYCN-3′UTR-2 siRNA and either control or let-7a miRNA mimic were harvested 48 hours post transfection. Control siRNA (Life Technologies #439846). LIN28B siRNAs: ORF1 (Life Technologies #4392420, ID:s52479), ORF2 (Life Technologies #4392420, ID:s52477), 5′UTR (GE Dharmacon Custom LIN28B-NM_001004317 Duplex siRNA, ON-TARGET Plus, sense: 5′-ACU GGA GAG AGG AGA GAA AUU-3′, antisense: 5′-UUU CUC UCC UCU CUC CAG UUU-3′), 3′UTR-1 (GE Dharmacon #J-028584-12-0020), 3′UTR-2 (GE Dharmacon Custom LIN28B-NM_001004317 Duplex siRNA, ON-TARGET Plus, sense: 5′-CAA CAG UGA UUG UGA GAA UUU-3′, antisense: 5′-AUU CUC ACA AUC ACU GUU GUU-3′). MYCN siRNAs: ORF-1 (Life Technologies #4392420, ID:s9135), ORF-2 (Life Technologies #4392420, ID:s9134). Control miRNA mimic (Life Technologies #4464059). let-7a miRNA mimic (Life Technologies #4464066, ID:MC10050), let-7a inhibitor (Life Technologies #4464084, ID:MH10050).
Powers J.T., Tsanov K.M., Pearson D.S., Roels F., Spina C.S., Ebright R., Seligson M., de Soysa Y., Cahan P., Theiβen J., Tu H.C., Han A., Kurek K.C., LaPier G.S., Osborne J.K., Ross S.J., Cesana M., Collins J.J., Berthold F, & Daley G.Q. (2016). Multiple mechanisms disrupt the let-7 microRNA family in neuroblastoma. Nature, 535(7611), 246-251.
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2

Reverse Transfection and Functional Assays

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BE(2)C and Kelly cells were reverse transfected using Lipofectamine 2000© (Life Technologies) into 6 well plates using the appropriate siRNA or miRNA mimics (described below). Cells were harvested at time-points described for analysis by western or qPCR. Growth assays were performed similarly, but in 96 well plates followed by time-point specific BrdU growth assay. Global let-7 target analysis: BE(2)C:MYCN-ORF cells co-transfected with control or MYCN-3′UTR-2 siRNA and either control or let-7a miRNA mimic were harvested 48 hours post transfection. Control siRNA (Life Technologies #439846). LIN28B siRNAs: ORF1 (Life Technologies #4392420, ID:s52479), ORF2 (Life Technologies #4392420, ID:s52477), 5′UTR (GE Dharmacon Custom LIN28B-NM_001004317 Duplex siRNA, ON-TARGET Plus, sense: 5′-ACU GGA GAG AGG AGA GAA AUU-3′, antisense: 5′-UUU CUC UCC UCU CUC CAG UUU-3′), 3′UTR-1 (GE Dharmacon #J-028584-12-0020), 3′UTR-2 (GE Dharmacon Custom LIN28B-NM_001004317 Duplex siRNA, ON-TARGET Plus, sense: 5′-CAA CAG UGA UUG UGA GAA UUU-3′, antisense: 5′-AUU CUC ACA AUC ACU GUU GUU-3′). MYCN siRNAs: ORF-1 (Life Technologies #4392420, ID:s9135), ORF-2 (Life Technologies #4392420, ID:s9134). Control miRNA mimic (Life Technologies #4464059). let-7a miRNA mimic (Life Technologies #4464066, ID:MC10050), let-7a inhibitor (Life Technologies #4464084, ID:MH10050).
Powers J.T., Tsanov K.M., Pearson D.S., Roels F., Spina C.S., Ebright R., Seligson M., de Soysa Y., Cahan P., Theiβen J., Tu H.C., Han A., Kurek K.C., LaPier G.S., Osborne J.K., Ross S.J., Cesana M., Collins J.J., Berthold F, & Daley G.Q. (2016). Multiple mechanisms disrupt the let-7 microRNA family in neuroblastoma. Nature, 535(7611), 246-251.
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3

Suv39H1 3'-UTR Luciferase Reporter Assay

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The 3′‑UTR of human Suv39H1 containing the potential miR‐125a‐5p binding sites was amplified by high fidelity PCR. Then, the amplified sequence was cloned into the XbaI site of the pGL3 control vector (Promega). The mutated putative miR‐125a‐5p binding site in the 3′‐UTR of Suv39H1 was generated using the QuickChange Site‐directed Mutagenesis kit (Stratagene) according to the manufacturer's protocol. The day before transfection, HEK293T cells were seeded in 24‐well plates (5 × 104 cells/well), then we transfected 500 ng Suv39H1 3′‐UTR‐pGL3 and 30 nmol/L (final concentration) miR‐125a‐5p mimics or control miRNA mimics (Ambion) into the cells with Lipofectamine 2000. Forty‐eight hours after transfection, luciferase activity was determined using dual‐luciferase reporter assay system (Promega) according to the manufacturer's protocol.
Cai M., Chen Q., Shen J., Lv C, & Cai L. (2018). Epigenetic silenced miR‐125a‐5p could be self‐activated through targeting Suv39H1 in gastric cancer. Journal of Cellular and Molecular Medicine, 22(10), 4721-4731.
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4

Regulation of Suv39H1 by miR-125a-5p

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The 3'-UTR of human Suv39H1 containing the potential miR-125a-5p was amplified by high-fidelity PCR. Then the amplified sequence was cloned into the XbaI site of the pGL3 control vector (Promega). The mutated putative miR-125a-5p binding site in the 3'-UTR of Suv39H1 was generated using the QuickChange Site-directed Mutagenesis kit (Stratagene) according to the manufacturer's protocol. The day before transfection, HEK293T cells were seeded in 24-well plates (5 × 104 cells well−1), then we transfected 500 ng Suv39H1 3'-UTR-pGL3 and 30 nM (final concentration) miR-125a-5p mimics or control miRNA mimics (Ambion) into the cells with Lipofectamine 2000. Forty-eight hours after transfection, luciferase activity was determined using dual-luciferase reporter assay system (Promega) according to the manufacturer's protocol.
Liu H., Ma Y., Liu C., Li P, & Yu T. (2018). Reduced miR-125a-5p level in non-small-cell lung cancer is associated with tumour progression. Open Biology, 8(10), 180118.
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5

Transfection of VSM Cells with miRNAs

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VSM cells (1 × 106) were harvested and electroporated with either miR-30c/e mimics (0.1 nmol) or miRNA mimic control (0.1 nmole) (Invitrogen) using the Amaxa Nucleofector system (Lonza) and the VSM cell-specific D33 program (Lonza Amaxa). After electroporation, cells were onto 4 × 60mm petri dishes and total RNA and protein were extracted 48 h and 72 h post-electroporation.
Liu Y.F., Spinelli A., Sun L.Y., Jiang M., Singer D.V., Ginnan R., Saddouk F.Z., Van Riper D, & Singer H.A. (2016). MicroRNA-30 inhibits neointimal hyperplasia by targeting Ca2+/calmodulin-dependent protein kinase IIδ (CaMKIIδ). Scientific Reports, 6, 26166.
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6

miR-20a Overexpression and Inhibition in A549 Cells

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Human miR-20a mimics (5'-UAAAGUGCUUAUAGUGCAGGUAG-3') and miR-20a inhibitor (5'-2'-O-methyl-CUACCUGCACUAUAAGCACUUUA-3') were synthesized by Invitrogen (Thermo Fisher Scientific, Inc.), and applied to overexpress or inhibit miR-20a expression in A549 cells using Lipofectamine® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.) following the manufacturer's protocol. The overexpression/inhibition was identified using RT-qPCR analysis after transfection (80 ng miR-20a mimics or inhibitor) for 48 h (23 (link)). miRNA mimic Control (5'-UUCUCCGAACGUGUCACGUUU-3'; Invitrogen; Thermo Fisher Scientific, Inc.) and miRNA inhibitor control (5'-2'-O-methyl-AAACGUGACACGUUCGGAGAA-3'; Invitrogen; Thermo Fisher Scientific, Inc.) were used as control after transfection (80 ng) for 48 h.
Wan K., Tu Z., Liu Z., Cai Y., Chen Y, & Ling C. (2021). Upregulated osteoprotegerin expression promotes lung cancer cell invasion by increasing miR-20a expression. Experimental and Therapeutic Medicine, 22(2), 846.
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