Mir 1 mimic

Manufactured by RiboBio

Sourced in China

The MiR-1 mimic is a laboratory reagent used in RNA research. It is a synthetic RNA molecule that mimics the function of the naturally occurring microRNA, miR-1. The core function of the MiR-1 mimic is to serve as a tool for studying the biological role and regulatory activities of miR-1 in cellular processes.

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Lab products found in correlation

6 protocols using mir 1 mimic

Isolation and Transfection of Rat Schwann Cells

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Primary SCs were isolated from the sciatic nerve of 1-day-old SD rats and further treated with anti-Thy1.1 antibody (Sigma, St Louis, MO) and rabbit complement (Invitrogen, Carlsbad, CA) to remove the fibroblasts as described previously21 (link). The final cell preparation consisted of 98% SCs, as determined by immunocytochemistry with SC marker anti-S100 (DAKO, Carpinteria, CA). A rat SC line (RSC96) was purchased from the American Type Culture Collection.
Primary SCs and RSC96 SCs were cultured in Dulbecco’s modified eagle medium (DMEM) containing 10% fetal bovine serum (FBS) in a humidified 5% CO₂ incubator at 37 °C. Primary SCs were passaged for no more than 3 times prior to use.
SC cultures were transfected with miR-1 mimic, miR-1 inhibitor, or BDNF siRNA (Ribobio, Guangzhou, China), respectively, using Lipofectamine RNAiMAX transfection reagent (Invitrogen) according to the manufacturer’s instructions.

Yi S., Yuan Y., Chen Q., Wang X., Gong L., Liu J., Gu X, & Li S. (2016). Regulation of Schwann cell proliferation and migration by miR-1 targeting brain-derived neurotrophic factor after peripheral nerve injury. Scientific Reports, 6, 29121.

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Modulation of Goat NK Cell Function by miR-1

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Goat NK cells were grown to logarithmic phase in 96-well plates with antibiotic-free medium the day before transfection. The miRNA transfection, including miR-1 mimic, mimic control (MC), miR-1 inhibitor, and inhibitor control (NC), was performed with Lipofectamine RNAiMAX (Life Technologies, USA) on cells of 50% confluence according to the manufacturer's protocol. The final concentrations of miR-1 mimic, miR-1 inhibitor, or their negative controls (RiBoBio, Guangzhou, China) were 100 nM. The effect of transfection was examined by quantitative RT-PCR and Western blot.

Qi X., Li Z., Li H., Wang T., Zhang Y, & Wang J. (2020). MicroRNA-1 Negatively Regulates Peripheral NK Cell Function via Tumor Necrosis Factor-Like Weak Inducer of Apoptosis (TWEAK) Signaling Pathways During PPRV Infection. Frontiers in Immunology, 10, 3066.

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Isolation and Manipulation of Neonatal Rat Ventricular Cardiomyocytes

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NRVCs were isolated from the hearts of 1- to 3-d old newborn SD rats as described previously16 (link). Fifty nM miR-1 mimic, 50 nM Hsp90aa1 siRNA and 100 nM miR-1 inhibitor (Ribobio, Guangzhou, China) were transfected into NRVCs by oligofectamine reagent (Invitrogen, Carlsbad, CA). NRVCs were infected with the recombinant Hsp90aa1 adenovirus and the control GFP adenovirus vector at a multiplicity of infection (MOI) of 10, respectively. NRVCs were serum-starved overnight prior to all experiments. Cell ischemic injury was induced by oxygen-glucose deprivation (OGD) treatment, and was followed with reoxygenation as previously described6 (link). Briefly, hypoxia was achieved by culturing NRVCs in serum- and glucose-free DMEM in a hypoxia chamber filled with 5% CO2 and 95% N2 at 37 °C for 4 hr. Then NRVCs were reoxygenated in DMEM containing 5% serum and normal glucose in a chamber filled with 5% CO2 and 95% O2 for 12 hr.

Zhu W.S., Guo W., Zhu J.N., Tang C.M., Fu Y.H., Lin Q.X., Tan N, & Shan Z.X. (2016). Hsp90aa1: a novel target gene of miR-1 in cardiac ischemia/reperfusion injury. Scientific Reports, 6, 24498.

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Transfection of miRNA Mimics and Inhibitors

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MiRNA mimics or inhibitors (mimic negative control: miR01101-1-5, miR-133a mimic: miR10000427-1-5, miR-1 mimic: miR10000416-1-5, miR-143 mimic: miR10000435-1-5, miR-145 mimic: miR10000437-1-5, miR-21 mimic: miR10000076-1-5, inhibitor negative control: miR02101-1-5, miR-133a inhibitor: miR20000427-1-5) were purchased from RiboBio (Guangzhou, China). Small RNA and plasmid transfections were performed with Lipofectamine® RNAiMAX Reagent and Lipofectamine® 3000 Reagent (Thermo Fisher Scientific), respectively, according to the manufacturer’s instructions.

Wei P., Xie Y., Abel P.W., Huang Y., Ma Q., Li L., Hao J., Wolff D.W., Wei T, & Tu Y. (2019). Transforming growth factor (TGF)-β1-induced miR-133a inhibits myofibroblast differentiation and pulmonary fibrosis. Cell Death & Disease, 10(9), 670.

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Dual-Luciferase Reporter Assay for miRNA Target Validation

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The 293T cell line used in this study was maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Waltham, United States) supplemented with 10% fetal bovine serum (Gibco), penicillin, and streptomycin in an incubator with 5% CO₂ at 37°C. Predicted binding sites were cloned and inserted into the pmirGLO vector (Promega, Madison, United States). For reporter assays, 150 ng of pmirGLO reporter vector and 50 nM miR-1 mimic (RiboBio, Guangzhou, China) were cotransfected into 293T cells using Lipofectamine 2000. No-mimic-treatment cells were used as a blank control, and cells carrying the pmirGLO-Hsp vector alone were used as the negative control. Firefly and Renilla luciferase activities were measured 48 h post-transfection with a Dual-Luciferase Reporter Assay System (Promega). First, 100 μl of luciferase assay reagent II was added to each well; firefly luciferase activities were measured. Subsequently, 100 μl of Stop&Glo reagent was added, and Renilla luciferase activities were measured. Firefly luciferase in the pmirGLO vector was used for normalization of Renilla luciferase expression. Treatments were assessed in triplicate, and transfections were repeated three times. Firefly luciferase activities were divided by Renilla luciferase activities for each experiment, providing the ratio.

Luo J., Ren Q., Liu W., Qiu X., Zhang G., Tan Y., Cao R., Yin H., Luo J., Li X, & Liu G. (2021). MicroRNA-1 Expression and Function in Hyalomma Anatolicum anatolicum (Acari: Ixodidae) Ticks. Frontiers in Physiology, 12, 596289.

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miR-1 Transcriptional Regulation Assay

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The 293T cell line used in this study was maintained in Dulbecco's modified Eagle's medium (DMEM) (Gibco, Waltham, USA) supplemented with 10% fetal bovine serum (Gibco), penicillin and streptomycin in an incubator with 5% CO 2 at 37°C. Predicted binding sites were cloned and inserted into the pmirGLO vector (Promega, Madison, USA). For reporter assays, 150 ng of pmirGLO reporter vector and 50 nM miR-1 mimic (RiboBio, Guangzhou, China) were cotransfected into 293T cells using Lipofectamine 2000. No-mimic-treatment cells were used as a blank control, and cells carrying the pmirGLO-Hsp vector alone were used as the negative control. Firefly and Renilla luciferase activities were measured 48 h posttransfection by Dual-Luciferase Reporter Assay System (Promega). First, 100 µL of luciferase assay reagent II was added to each well, firefly luciferase activities were measured, and 100 µL Stop&Glo reagent was then added. Renilla luciferase activities were then measured. Firefly luciferase in the pmirGLO vector was used for normalization of Renilla luciferase expression. Treatments were assessed in triplicate, and transfections were repeated three times. Firefly luciferase activities were divided by Renilla luciferase activities for each experiment, providing the ratio.

Luo J., Ren Q., Liu W., Qiu X., Zhang G., Tan Y., Cao R., Yin H., Luo J., Li X., & Liu G. (2020). MicroRNA-1 promotes the development of and prolongs engorgement time in Hyalomma anatolicum anatolicum (Acari: Ixodidae) ticks.

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