Mirna nc

Manufactured by GenePharma

Sourced in China

The MiRNA NC is a laboratory instrument designed for the detection and analysis of microRNA (miRNA) expression. It functions as a platform for the quantification and profiling of miRNA molecules in biological samples.

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34 protocols using mirna nc

miR-1246 and -1290 Modulation

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The mimics and inhibitors of miR-1246 and -1290 as well as the corresponding negative control (NC) miRNAs were obtained from GenePharma (Shanghai, China). The siRNAs for silencing EFL5 and SP1 and the respective scrambled siRNAs as negative controls were designed and synthesized by GenePharma. The ELF5- and SP1-overexpressing plasmids (pcDNA3.0-ELF5 and pcDNA3.0-SP1) as well as the empty pcDNA3.0 plasmids were purchased from GenePharma. Cell transfections with miRNAs, siRNAs, or plasmids were performed using Lipofectamine 2000 reagent (Invitrogen, Waltham, United States) in accordance with the manufacturer’s protocols.

Huang Y., Chen L., Feng Z., Chen W., Yan S., Yang R., Xiao J., Gao J., Zhang D, & Ke X. (2021). EPC-Derived Exosomal miR-1246 and miR-1290 Regulate Phenotypic Changes of Fibroblasts to Endothelial Cells to Exert Protective Effects on Myocardial Infarction by Targeting ELF5 and SP1. Frontiers in Cell and Developmental Biology, 9, 647763.

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miRNA Inhibition Transfection Protocol

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The cells were transfected with an inhibitory oligonucleotide targeting miR-29c or miR-19b (miRNA inhibitor, 100 nM) or with nonspecific control (NC) miRNAs (GenePharma, Shanghai, China) using Lipofectamine™ 2000 reagent (Invitrogen) according to the manufacturer's instructions. Five samples were tested in per group.

Wu Y., Xu J., Xu J., Cheng J., Jiao D., Zhou C., Dai Y, & Chen Q. (2017). Lower Serum Levels of miR-29c-3p and miR-19b-3p as Biomarkers for Alzheimer's Disease. The Tohoku journal of experimental medicine, 242(2).

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Biotinylated miRNA Pulldown Assay

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Biotinylated miR-10b and its NC miRNA (both of which were biotin-labeled at the 3′ end) were purchased from GenePharma (Shanghai, China). hESC-CMs were transfected with NC miRNA and miR-10b using Lipofectamine RNAiMAX. Twenty-four hours after transfection, the cells were lysed with lysis buffer (20 mM Tris-HCl [pH 7.5], 100 mM KCl, 5 mM MgCl, 0.3% IGEPAL CA-630), and the lysate (approximately 500 μL) was mixed with prewashed streptavidin beads overnight at 4°C. The remaining lysate was stored at −80°C as an input. The next day, the mixture was washed with lysis buffer, and RNA was extracted with TRIzol reagent (Invitrogen). Then, quantitative real-time PCR was performed according to the standard protocol.

Xie Y., Wang Q., Gao N., Wu F., Lan F., Zhang F., Jin L., Huang Z., Ge J., Wang H, & Wang Y. (2019). MircroRNA-10b Promotes Human Embryonic Stem Cell-Derived Cardiomyocyte Proliferation via Novel Target Gene LATS1. Molecular Therapy. Nucleic Acids, 19, 437-445.

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miR-30a Mimic Transfection in Cells

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miR-30a mimics (20 nmol/l) and negative control (NC) miRNA (20 nmol/l) were acquired from Shanghai GenePharma Co., Ltd. (Shanghai, China). The miR-30a mimics sequence was 5′-UGUAAACAUCCUCGACUGGAAG-3′. The NC miRNA sequence was 5′-ACAUUUGUAGGAGCUGACCGGC-3′. Lipofectamine 2000^® (Invitrogen; Thermo Fisher Scientific, Inc.) was subsequently utilized to perform transient transfection following incubation at 37°C for 6 h. Transfected cells were collected and purified after 48 h incubation.

Guo H, & Zhang L. (2019). MicroRNA-30a suppresses papillary thyroid cancer cell proliferation, migration and invasion by directly targeting E2F7. Experimental and Therapeutic Medicine, 18(1), 209-215.

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Cardiomyocyte Differentiation and Transfection

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Hearts were dissected from E8.5–9.5 mouse embryos, washed in PBS without Ca²⁺, and digested using 0.04% Trypsin and 0.05% Collagenase IV. After incubation at 37 °C for 20 min (E8.5 embryonic hearts) or 30 min (E9.5 embryonic hearts), cells were gently pipetted and transferred to DMEM containing 10% fetal bovine serum for termination. Cells were plated on matrigel (1:60, BD) coated plates in the in vitro CMs culture medium (IVCC), with 4 ng/mL zbFGF. The in vitro CMs culture medium (IVCC) is composed of DMEM/F12 (Gibco) medium with 2% B27 (Gibco), 1% N2 (Gibco), 1% Glutamax (Gibco), 1% non-essential amino acids (NEAA; Gibco), 100 units/mL penicillin and 100 µg/mL streptomycin (Gibco). 50 µg/mL 2-phospho-l-ascorbic acid (Vitamin C, Sigma), 0.001% Progesterone (Sigma), 1% MEM Vitamin (Sigma, M6875), 0.1% trace element B (Corning), and 0.1% trace element C (Corning).
After 24 h culture, CMs were transfected with 50 nmol/L miRNA mimics (miRNA-1-3p, miR-541-5p and NC-miRNA (GenePharma) using Lipofectamine^® 2000 (Thermo Fisher) following the manufacturer’s instructions. The same transfection protocol was used for SEVC4–10, mouse Yolk sac dissociated cells, human iPSC derived ECs and mESCs.

Chen X., Wang L., Huang R., Qiu H., Wang P., Wu D., Zhu Y., Ming J., Wang Y., Wang J, & Na J. (2018). Dgcr8 deletion in the primitive heart uncovered novel microRNA regulating the balance of cardiac-vascular gene program. Protein & Cell, 10(5), 327-346.

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Modulating miR-93-5p Expression in SCCHN

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SCCHN cells were transfected with the miR-93-5p mimics, the miR-93-5p inhibitors, or with negative control (NC) miRNA (Genepharma, Suzhou, China) using si-Mate^TM (Genepharma) according to the manufacturer's protocol. Additionally, to establish a cell line that stably expressed miR-93-5p, Tu686 cells were infected with lentivirus expressing miR-93-5p (GeneCopoeia Inc., MD, USA), and the infected cells were selected in the medium containing 3 μg/mL puromycin.

Zhang S., He Y., Liu C., Li G., Lu S., Jing Q., Chen X., Ma H., Zhang D., Wang Y., Huang D., Tan P., Chen J., Zhang X., Liu Y, & Qiu Y. (2020). miR-93-5p enhances migration and invasion by targeting RGMB in squamous cell carcinoma of the head and neck. Journal of Cancer, 11(13), 3871-3881.

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Modulating miR-144-3p Expression

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For the manual alteration of miR-144-3p expression, negative control miRNA (NC miRNA), miRNA-mimics and miRNA-inhibitor (GenePharma, China) were transfected into SH-SY5Y cells using Lipofectamine 2000 Reagent (Invitrogen, USA) according to the manufacturer’s instructions.

Li K., Zhang J., Ji C, & Wang L. (2016). MiR-144-3p and Its Target Gene β-Amyloid Precursor Protein Regulate 1-Methyl-4-Phenyl-1,2-3,6-Tetrahydropyridine-Induced Mitochondrial Dysfunction. Molecules and Cells, 39(7), 543-549.

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Transfection of PLAC2 and XiaP in Cancer Cells

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pcDNA3-PLAC2 and pcDNA3-XiaP vectors, NC siRNA, lncRNA PLAC2 siRNA, miR-96 mimic, and NC miRNA were obtained from GenePharma (Shanghai, China). SNU-475 and SNU-387 cells were harvested at 70–80% confluence, and 12 nM lncRNA PLAC2 or XiaP expression vector, empty pcDNA3 vector (negative control, NC), 50 nM lncRNA PLAC2 siRNA, or negative control siRNA (negative control, NC) was transfected using Lipofectamine 2000 (Cat# 11668019, Invitrogen, Shanghai, China).

Wang H., Zhang X., Chen X., Zhang S., Yun Z., Gao Q., Sheng H, & Wang J. (2022). Long non-coding RNA placenta‑specific protein 2 regulates the chemosensitivity of cancer cells to cisplatin in hepatocellular carcinoma (HCC) by sponging microRNA-96 to upregulate X-linked inhibitor of apoptosis protein. Bioengineered, 13(4), 10765-10773.

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miR-124 Mimics in Esophageal Cancer

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The miR-124 mimics and the negative control (NC) miRNA were synthesized by GenePharma (Shanghai, China) and transfected into TE-1 cells to a final oligonucleotide concentration of 30 nM. All cell transfections were performed with Lipofectamine TM 2000 reagent (Invitrogen) according to the manufacturer instructions. For each treatment group, the experiments were performed in triplicate. Cells were collected for further experiments 48 h post-transfection.

Zhang Y.H., Wang Q.Q., Li H., Ye T., Gao F, & Liu Y.C. (2016). miR-124 radiosensitizes human esophageal cancer cell TE-1 by targeting CDK4. Genetics and molecular research : GMR, 15(2).

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Targeting circ_0062020 in Prostate Cancer

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Short hairpin RNAs (shRNAs) stably knocked out circ_0062020 (sh-circ_0062020), small interfering RNAs (siRNAs) specifically inhibited circ_0062020 (si-circ_0062020) expression, miR-615-5p inhibition (miR-615-5p inhibitor), miR-615-5p overexpression (miR-615-5p mimic), TRIP13 overexpression (pc-TRIP13), and the corresponding negative controls (sh-NC, si-NC, inhibitor NC, miRNA NC, pc-NC) were synthesized and provided by GenePharma (Shanghai, China). The transfection was performed by Lipofectamine2000 (Invitrogen) to DU145 and LNCaP cells according to the manufacturer’s protocol.

Li H., Zhi Y., Ma C., Shen Q., Sun F, & Cai C. (2020). Circ_0062020 Knockdown Strengthens the Radiosensitivity of Prostate Cancer Cells. Cancer Management and Research, 12, 11701-11712.

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