In mir nc

Manufactured by GenePharma

Sourced in China

In-miR-NC is a laboratory reagent used in molecular biology experiments. It serves as a negative control for microRNA (miRNA) studies. The product is designed to monitor the effects of miRNA regulation without introducing any specific miRNA target.

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MiR-NC (532 citations) MiR-in (6 citations) In-NC (4 citations)

28 protocols using in mir nc

Gastric Carcinoma Cell Lines Transfection

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Two human gastric carcinoma cell lines MGC-803 (CQ80145) and AGS (H007) and human stomach normal epithelial cell lines GES-1 (H054) were purchased from ChuanQiu Biotechnology (Shanghai, China). All cells were cultivated in Dulbecco’s modified Eagle’s medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific) and 1% penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA) in an incubator with the conditions of 37°C and 5% CO₂.
Small interfering RNA (siRNA) targeting HOTAIR (si-HOTAIR#1, si-HOTAIR#2, si-HOTAIR#3) and negative control (si-NC), miR-618 mimics (miR-618) and negative control (miR-NC), miR-618 inhibitor (in-miR-618) and negative control (in-miR-NC), HOTAIT overexpression plasmid (HOTAIR) and KLF12 overexpression vector (KLF12) were all obtained from GenePharma (Shanghai, China). The transfection was performed using Lipofectamine 2000 (Invitrogen) according to the reference.

Xun J., Wang C., Yao J., Gao B, & Zhang L. (2019). Long Non-Coding RNA HOTAIR Modulates KLF12 to Regulate Gastric Cancer Progression via PI3K/ATK Signaling Pathway by Sponging miR-618. OncoTargets and therapy, 12, 10323-10334.

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Regulating TUG1 and Rab10 in Acute Myeloid Leukemia

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Small interfering RNAs against TUG1 (si-TUG1#1, si-TUG1#2 and si-TUG1#3) and the scrambled negative control si-NC, miR-193a-5p mimics (miR-193a-5p), miR-193a-5p inhibitor (in-miR-193a-5p) and their matched negative controls (miR-NC, in-miR-NC), si-Rab10 and its negative control si-NC, as well as the pcDNA-TUG1 (TUG1) and pcDNA-Rab10 (Rab10), were synthesized by GenePharma Co., ltd (Shanghai, China). The pcDNA plasmid and transfection reagent Lipofectamine 3000 were purchased from Thermo Fisher Scientific (Waltham, MA, USA). The aforementioned oligonucleotides or plasmids were transfected into HL-60 and NB4 cells. In brief, cells were seeded in 24-well plates for 24 hrs. When cell confluence reached 70%, 100 μL RPMI1640 medium without serum mixed with constructed oligonucleotides (40 nM) + 2 μL Hiperfect transfection reagents QIAGEN (Dusseldorf, Germany) or plasmids (2 μg) + 3 μL Lipofectamine were incubated at 37°C for 10 mins. Then, the complexes were drop-wise added onto the cells in 24-well plates gently. Forty-eight-hour later, transfected HL-60 and NB4 cells were subjected for subsequent assays.

Li Q, & Wang J. (2020). LncRNA TUG1 Regulates Cell Viability and Death by Regulating miR-193a-5p/Rab10 Axis in Acute Myeloid Leukemia. OncoTargets and therapy, 13, 1289-1301.

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Regulation of ox-LDL-induced HUVEC dysfunction

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NGR1 was purchased from Azelasi Biotechnology (China) and diluted according to the manufacturer's instructions. After treatment with 30 µM NGR1 for 24 h, HUVECs were treated with 50 mg/L ox-LDL (Bioss, China) for 24 h in serum-free medium. miR-221-3p mimic and inhibitor (miR-221-3p and in-miR-221-3p) or their negative controls (miR-NC and in-miR-NC), TLR4 overexpression plasmid (TLR4), and its negative control (pcDNA) were purchased from GenePharma (China). Lipofectamine 2000 (Invitrogen, USA) was used to transfect these plasmids into HUVECs. After transfection for 24 h, HUVECs were treated with ox-LDL or NGR1.

Zhu L., Gong X., Gong J., Xuan Y., Fu T., Ni S., Xu L, & Ji N. (2020). Notoginsenoside R1 upregulates miR-221-3p expression to alleviate ox-LDL-induced apoptosis, inflammation, and oxidative stress by inhibiting the TLR4/NF-κB pathway in HUVECs. Brazilian Journal of Medical and Biological Research, 53(6), e9346.

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Modulating circRASGRF2 Expression in HCC

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siRNAs that were used to knock down endogenous circRASGRF2 expression (si-circRASGRF2) and si-NC were synthesized by RiboBio (Guangzhou, China). A miR-1224 mimic, miRNA negative control (miR-NC), miR-1224 inhibitor (in-miR-1224), and negative control inhibitor (inmiR- NC) were acquired from GenePharma (Shanghai, China). For the overexpression of circRASGRF2, the sequences of circRASGRF2 were inserted into vector pcDNA3.1 to construct pcDNA3.1/circRASGRF2 (termed circRASGRF2), and empty vector was considered as a negative control in this study. HCC cells were seeded in six-well plates, and cell transfection was conducted using the Lipofectamine 2000 reagent (Invitrogen). Cells transfected with the aforementioned oligonucleotide(s) or plasmid(s) were harvested and used in subsequent experiments.

Wu D., Xia A., Fan T, & Li G. (2020). circRASGRF2 functions as an oncogenic gene in hepatocellular carcinoma by acting as a miR-1224 sponge. Molecular Therapy. Nucleic Acids, 23, 13-26.

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Silencing LINC00514 and URGCP in Osteosarcoma

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Three siRNAs targeting LINC00514 (si-LINC00514#1, si-LINC00514#2, and si-LINC00514#3) were designed to silence endogenous LINC00514. si-URGCP was used to suppress URGCP expression. All these siRNAs and si-NC were designed and chemically synthesized by Guangzhou RiboBio Co., Ltd. (RiboBio, Guangzhou, China). MiR-708 mimic, miR-NC, in-miR-708, and in-miR-NC were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China). The URGCP overexpression plasmid pCDNA3.1-URGCP (pc-URGCP) and an empty pcDNA3.1 plasmid were obtained from GeneChem Co., Ltd. (Shanghai, China). The OS cells were seeded into 6-well plates 1 day before transfection and transfected with the aforementioned molecular products using Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA).

Yu D., Xu X., Li S, & Zhang K. (2020). LINC00514 drives osteosarcoma progression through sponging microRNA-708 and consequently increases URGCP expression. Aging (Albany NY), 12(8), 6793-6807.

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Gastric Cancer Cell Lines Transfection

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Several GC cell lines, including N87, HGC27, and AGS, were bought from ATCC. Human normal gastric cell (GES‐1) and GC cell line (MKN‐45) were commercially gained from Chinese Academy of Sciences (Shanghai, China). The RPMI‐1640 medium (Gibco) supplemented with 10% fetal bovine serum (FBS; Gibco) was used to culture cells under a moist environment with 5% CO₂ at 37°C.
Small interfering RNAs (siRNAs) targeting hsa_circ_0043691 (si‐hsa_circ_0043691), miR‐1294 mimic (miR‐1294), inhibitor (in‐miR‐1294) and their relative control (si‐NC, miR‐NC and in‐miR‐NC) were constituted from GenePharma. Plasmid of pcDNA3.1‐PBX3 (PBX3), and pcDNA3.1 (pcDNA) were composed and obtained from Origene. Transient transfection of the above‐mentioned oligonucleotides or vectors were executed by Lipofectamine 3000 (Thermo Fisher).

Wu J., Gong P, & Jiang Z. (2022). Knockdown of hsa_circ_0043691 restrains the progression of gastric cancer by decoying miR‐1294 to target pre‐leukemia transcription factor 3. Journal of Clinical Laboratory Analysis, 36(12), e24733.

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Manipulating ANO1 expression in RAW264.7 cells

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The ANO1 sequences were cloned into pcDNA3.1 vector (Thermo Fisher, Wilmington, DE, USA) to generate the overexpression vector of ANO1, with empty vector as control (pcDNA). The short interfering RNA (siRNA) for ANO1 (si-ANO1, 5′-AAGUAUAGUCCAUACUUGCAU-3′), siRNA negative control (si-NC, 5′-UUCUCCGAACGUGUCACGU-3′), miR-106a mimic (5′-CAAAGUGCUAACAGUGCAGGUAG-3′), mimic negative control (miR-NC, 5′-CGAUCGCAUCAGCAUCGAUUGC-3′), miR-106a inhibitor (in-miR-106a, 5′-UAGAACUCAAAAAGCUACCUG-3′), and inhibitor negative control (in-miR-NC, 5′-CUAACGCAUGC ACAGUCGUACG-3′) were provided by GenePharma (Shanghai, China). Transfection was conducted in RAW264.7 cells via Lipofectamine 3000 (Thermo Fisher) for 24 h.

Heng J., Wu D., Lu S, & Zhao Y. (2020). miR-106a Targets Anoctamin 1 (ANO1) to Regulate Lipopolysaccharide (LPS)-Induced Inflammatory Response in Macrophages. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 26, e922479-1-e922479-11.

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Investigating circRANBP17/miR-27b-3p/KDM1A Axis

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NB cell-lines SK-N-AS (CRL-2137), SK-N-SH (HTB-11), BE(2)-C (CRL-2268), IMR-32 (CCL-127) and human umbilical vein endothelial cell-line HUVEC (CRL-1730) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). All cells were cultured in DMEM (Invitrogen, Carlsbad, USA) containing 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Rockville, MD, USA) at 37°C in an incubator with 5% CO₂. Small interference RNA (siRNA) targeting circRANBP17 (si-circRANBP17, 5′-GCCCAGTGTTTGCCAAACCTT-3′) and its mock (si-NC, 5′-CCTCTACCTGTCGCTGAGCTGTAAT-3′), miR-27b-3p mimics (miR-27b-3p, 5′-UUCACAGUGGCUAAGUUCUGC-3′) and its matched control (miR-NC, 5′-UUUGUACUACACAAAAGUACUG-3′), miR-27b-3p inhibitor (in-miR-27b-3p, 5′-GCAGAACUUAGCCACUGUGAA-3′) and its matched control (in-miR-NC, 5′-CAGUACUUUUGUGUAGUACAAA-3′) and KDM1A overexpression vector (KDM1A) and empty plasmid (pcDNA) were obtained from GenePharma (Shanghai, China). Lipofectamine 2000 Reagent (Invitrogen) was used for transfection process following the guidebook. The primers used to amplify the full length of coding sequence of KDM1A were as follows: 5′-GGAATTCCATGTTATCTGGGAAGAAGGC-3′ and 5′-CCGCTCGAGCGGTCACATGCTTGGGGACT-3′.

Zhao L., Fan J., Zhang C., Zhang Z, & Dong J. (2023). CircRANBP17 modulated KDM1A to regulate neuroblastoma progression by sponging miR-27b-3p. Open Medicine, 18(1), 20230672.

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Overexpression and Silencing of TUG1 in Podocytes

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To over-express TUG1, the cDNA sequences of TUG1 were cloned in pcDNA3.1 empty vector (pcDNA; Invitrogen), termed as pcDNA3.1-TUG1 (TUG1). siRNA against TUG1(si-TUG1), siRNA against E2F3 (si-E2F3) and corresponding scrambled negative control (si-NC), miR-27a-3p mimic (miR-27a-3p), miR-27a-3p inhibitor (in-miR-27a-3p), and scrambled negative controls (miR-NC, in-miR-NC) were purchased from GenePharma (Shanghai, China). Transient transfection of the indicated oligonucleotides and plasmids into treated podocytes was implemented by Lipofectamine 2000 reagents (Invitrogen) based on the operation manual.

Li Y., Huang D., Zheng L., Cao H., Gao Y., Yang Y, & Fan Z. (2019). Retracted Article: Long non-coding RNA TUG1 alleviates high glucose induced podocyte inflammation, fibrosis and apoptosis in diabetic nephropathy via targeting the miR-27a-3p/E2F3 axis. RSC Advances, 9(64), 37620-37629.

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circZNF532 regulation in hRMECs

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circZNF532 small interfering RNA (si-circZNF532) and the control (si-NC), miR-1243 mimics (miR-1243) and the control (miR-NC), miR-1243 inhibitor (in-miR-1243) and the control (in-miR-NC), CARM1 overexpression vector (CARM1) and the control (pcDNA) were purchased from GenePharma (Shanghai, China). Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) was employed for cell transfection when hRMECs reached ~ 80% confluence.

Wang T., Li C., Shi M., Zhou S., Chen J, & Wang F. (2022). Circular RNA circZNF532 facilitates angiogenesis and inflammation in diabetic retinopathy via regulating miR-1243/CARM1 axis. Diabetology & Metabolic Syndrome, 14, 14.

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