Mir 149 3p mimic

Manufactured by GenePharma

Sourced in China

MiR-149-3p mimics are small, chemically modified RNA molecules that mimic the function of the naturally occurring microRNA miR-149-3p. MicroRNAs are important regulators of gene expression, and miR-149-3p has been associated with various biological processes. The mimics are designed to be used in research applications to study the role and function of miR-149-3p.

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4 protocols using mir 149 3p mimic

Investigating LINC00665 and miR-149-3p in Gastric Cancer

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Human GC cells (AGS, SGC-7901, HGC27, MGC-803, MKN-45, and BGC-823) and normal gastric epithelial cells (GES-1) were bought from American Type Culture Collection (ATCC, USA) and maintained in RPMI1640 medium (Gibco, NY, USA) supplemented with 10% fetal bovine serum (FBS; Gibco), 100 U/mL penicillin (Gibco), and 100 mg/mL streptomycin (Gibco) at 37 °C in a humidified atmosphere with 5% CO₂.
For the knockdown of LINC00665, sh-LINC00665 #1/2/3 were obtained from Ribo Biotech (Guangzhou, China). miR-149-3p mimics, miR-149-3p inhibitors, and overexpression RNF2 vector (pCDNA3.1-RNF2) were obtained from GenePharma (Shanghai, China). GC cells transfected with mimics or plasmid vector by Lipofectamine 2000 (Invitrogen) according to the protocol.

Qi H., Xiao Z, & Wang Y. (2019). Long non-coding RNA LINC00665 gastric cancer tumorigenesis by regulation miR-149-3p/RNF2 axis. OncoTargets and therapy, 12, 6981-6990.

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Transfection of miR-149-3p and p53 modulators

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miR-149-3p mimics, inhibitors, siPDK2, sip53, and their corresponding NC oligonucleotide sequences were synthesized by GenePharma (Shanghai, China). Flag-p53 plasmid was kindly provided by Dr. Lu (Hua Lu, Fudan University, Shanghai, China). Transfection was performed with Lipofectamine 3000 (Invitrogen, CA, USA) at a final concentration of 50 nmol/L (mimics and siRNAs) or 100 nmol/L (inhibitors). Cells were harvested for assays 24 or 48 h after transfection. The siRNA, mimic, and inhibitor sequences are shown in Supplementary Table 2.

Liang Y., Hou L., Li L., Li L., Zhu L., Wang Y., Huang X., Hou Y., Zhu D., Zou H., Gu Y., Weng X., Wang Y., Li Y., Wu T., Yao M., Gross I., Gaiddon C., Luo M., Wang J, & Meng X. (2019). Dichloroacetate restores colorectal cancer chemosensitivity through the p53/miR-149-3p/PDK2-mediated glucose metabolic pathway. Oncogene, 39(2), 469-485.

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Regulation of Colorectal Cancer by HOXA11-AS

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A small interfering RNA (siRNA) against HOXA11-AS (HOXA11-AS-siRNA), NC siRNA, miR-149-3p inhibitor, and miR-149-3p mimic were obtained from Gene Pharma (Shanghai, China). The HCT116 human CRC cells (2 × 10⁵ cells) were transfected with these constructs at a final concentration of 25 nmol/L using Lipofectamine 2000 Reagent (Life Technologies, Carlsbad, CA, USA). Cells were transfected with the pcDNA-HOXA11-AS constructs at a final concentration of 1 μg/μL according to the manufacturer's protocol. After transfection for 48 h, total RNA was isolated from the harvested cells using TRIzol reagent (Invitrogen). Empty pEX3 vector and scrambled sequences of the miR-149-3p mimics, miR-149-3p inhibitor, or siRNAs were used as NCs.

Chen D., Zhang M., Ruan J., Li X., Wang S., Cheng X., Zhao H., Zeng Y., Liu J., He K, & Zhao P. (2020). The long non-coding RNA HOXA11-AS promotes epithelial mesenchymal transition by sponging miR-149-3p in Colorectal Cancer. Journal of Cancer, 11(20), 6050-6058.

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Establishment of Stable Transfected Cells

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For the establishment of stably transfected cells, lentiviruses carrying ZEB1-AS1 and ZEB1-AS1-sh-RNA vectors were constructed (Wanleibio, Shenyang, China) and respectively transfected into SGC-7901 and MGC-803 cells according to the manufacturer’s protocol. Following 48 h of transfection, cells were selected with 10 μg/mL puromycin, measured through a fluorescent inverted microscope, with qPCR performed to determine the transfection efficiency (Additional file 1: Fig. S1). The miR-149-3p mimic and the miR-149-3p negative control (miR-NC) elements (GenePharma Corporation, Shanghai, China) were transiently and separately transfected into the same cell lines using Lipofectamine TM 3000 transfection reagent (Invitrogen, Carlsbad, CA, USA). Cells and transfection complexes were co-incubated for 5 h. Transfection efficiency in these cells was also validated by qPCR. The sequences involved in the study are detailed in Additional file 2: Table S1 and Additional file 3.

Ma M.H., An J.X., Zhang C., Liu J., Liang Y., Zhang C.D., Zhang Z, & Dai D.Q. (2019). ZEB1-AS1 initiates a miRNA-mediated ceRNA network to facilitate gastric cancer progression. Cancer Cell International, 19, 27.

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