miR-1 mimics, scramble miRNA negative control (miR-NC) and miR-1 inhibitors were synthesized by Shanghai GenePharma Co., Ltd.. The sequences were as follows: miR-1 mimic, 5′-UGGAAUGUAAAGAAGUAUG UAU-3′ (sense) and 5′-ACAUACUUCUUUACAUUCCAUU-3′ (antisense); miR-NC, 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); miR-1 inhibitor, 5′-UUCAGUUAUCACAGUACU GUA-3′; inhibitor NC, 5′-CAGUACUUUUGUGUAGUA CAA-3′. SGC7901, SGC7901/ADM and SGC7901/VCR cells were transfected with 100 nM miR-1 mimics or miR-1 inhibitor for miR-1 overexpression and knockdown, respectively. miR-NC (100 nM) and inhibitor NC (100 nM) were used as negative controls. Transfection was conducted using Lipofectamine 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. Six hours post-transfection, the medium was changed with fresh medium. At 48 h post-transfection, the cells were used in subsequent experiments.
Mir 1 mimic
Manufactured by GenePharma
Sourced in China
The MiR-1 mimics is a laboratory product designed for molecular biology research. It is a synthetic molecule that mimics the function of the naturally occurring microRNA-1 (miR-1). The core function of the MiR-1 mimics is to facilitate the study of miR-1 and its regulatory roles in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (3 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Mir nc, by GenePharma (2 mentions)
Inhibitor nc, by GenePharma (1 mentions)
Pcdna3, by Thermo Fisher Scientific (1 mentions)
Si nc, by GenePharma (1 mentions)
Rpmi 1640, by Lonza (1 mentions)
Heepic, by ScienCell (1 mentions)
Si hotair, by GenePharma (1 mentions)
Pcr system, by Thermo Fisher Scientific (1 mentions)
Taqman real time polymerase chain reaction, by Thermo Fisher Scientific (1 mentions)
Abi prism 7900ht, by Thermo Fisher Scientific (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
Sw480, by American Type Culture Collection (1 mentions)
Sw620, by American Type Culture Collection (1 mentions)
8 protocols using mir 1 mimic
1
miR-1 Overexpression and Knockdown in Gastric Cancer
2
Synthetic Oligonucleotides for miR-1 and SNAP25 Regulation
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MiR-1 mimics and AMO-1 for rats were synthesized by Shanghai GenePharma Co., Ltd. (Shanghai, China). AMO-1 contains 2’-O-methyl modifications. In addition, scrambled RNA (mis-miR-1) was used as a negative control. Snap25-masking antisense oligodeoxynucleotides (ODNs) were synthesized by Shanghai Sangon Biotech Co., Ltd., China, and these ODNs complemented the position of 408–430 containing the binding sites of miR-1 located at position 413–420 in the 3’ UTR of Snap25. The nucleotides or deoxynucleotides at both ends of the antisense molecules were locked by a methylene bridge connecting the 2’-O and 4’-C atoms. The sequences of these synthesized oligonucleotides are shown in Table 1 .
The sequences of the synthesized oligonucleotides
| miR-1 mimics | sense: 5’-UGGAAUGUAAAGAAGUGUGUAUGU-3′ |
| AMO-1 | 5’-ACCUUACAUUUCUUCACACAUACA-3’ |
| mis-miR-1 | sense: 5’-UUCUCCGAACGUGUCACGUAA-3′ |
| Snap25-ODN | 5’GUAGCUCUGUGGAAUGUCACAG-3’ |
3
Transfection of miR-1 Mimics and Inhibitors
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miR-1mimics, miR-1 inhibitor, NC and inhibitor NC were designed and synthesized by GenePharma (Shanghai, China). Solutions were dissolved in DEPC water to reach a final concentration of 20 µM. In a six-well plate, approximately 5–6 × 105 cells in logarithmic growth phase in medium containing serum and double antibody were added to each well. miR-1 mimics, NC and serum-free 1640 medium were added to TE-1 and KYSE410 cells to a final concentration of 50 nM. miR-1 inhibitor, inhibitor NC and serum-free medium were added to TE-1 and KYSE410 cells to a final concentration of 150 nM. For each group, 12 μl of HiPerFect transfection reagent was added to the samples. After incubation at room temperature for 5–10 min, the mixture was added to the cells, and the cells were placed in an incubator.
4
miR-1 Gain/Loss-of-Function in KYSE-150 Cells
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The miR-1 gain-of-function experiments using KYSE-150 cells were performed using miR-1 mimics (100 nM) purchased from Shanghai GenePharma Co., Ltd., (Shanghai, China) and its negative control (NC, 100 nM). The loss-of-function experiments using KYSE-150 cells were performed using miR-1 inhibitor (100 nM) and its NC (100 nM). The miR-1 inhibitor was 2′-O-methyl oligoribonucleotides, purchased from GenePharma. For each experiment, we included a negative control. The cells were transfected using Lipofectamine™ 2000 (Invitrogen) in Opti-MEM (Gibco) according to the manufacturer's instructions. The transfection efficiency was confirmed by the RT-qPCR detection of miR-1 expression.
5
Role of HOTAIR and miR-1 in ESCC
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Six ESCC cell lines (KYSE30, KYSE140, KYSE150, KYSE180, KYSE410, and KYSE510) were purchased from the German Culture Collection (DSMZ, Braunschweig, Germany). Human normal esophageal epithelium cells (HEEpiC) were obtained from ScienCell (Carlsbad, CA). These cells were grown in RPMI 1640 (Biowhittaker, Walkersville, MD) supplemented with 10% fetal bovine serum (HyClone, Logan, UT) at 37°C under 5% CO2 and saturated moisture.
For investigating the role of HOTAIR, KYSE30 and KYSE510 cells were transfected with either siRNAs targeting HOTAIR (si-HOTAIR) or scrambled negative controls (si-NC; GenePharma, Shanghai, China). For investigating the relationship between HOTAIR and miR-1, HOTAIR cDNA and miR-1 mimics (GenePharma) were cloned into the mammalian expression vector pcDNA3.1 (Invitrogen, Carlsbad, CA) individually or in combination, and then the vectors were transfected into cells. For the function study of CCND1, KYSE30 and KYSE510 cells were transfected with either si-CCND1 or si-NC. All transfection reactions were performed by using Lipofectamine 2000 (Invitrogen) following the manufacturer's instructions.
For investigating the role of HOTAIR, KYSE30 and KYSE510 cells were transfected with either siRNAs targeting HOTAIR (si-HOTAIR) or scrambled negative controls (si-NC; GenePharma, Shanghai, China). For investigating the relationship between HOTAIR and miR-1, HOTAIR cDNA and miR-1 mimics (GenePharma) were cloned into the mammalian expression vector pcDNA3.1 (Invitrogen, Carlsbad, CA) individually or in combination, and then the vectors were transfected into cells. For the function study of CCND1, KYSE30 and KYSE510 cells were transfected with either si-CCND1 or si-NC. All transfection reactions were performed by using Lipofectamine 2000 (Invitrogen) following the manufacturer's instructions.
6
Transient Transfection of miR-1 Mimics
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The cells were transiently transfected with 50 nM miR-1 mimics or the 50 nM negative control (Genepharma, Inc., Sunnyvale, CA, USA) using Lipofectamine™ 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) diluted in Opti-MEM (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. The negative control was a scrambled oligonucleotide not encoding any known miRNA. The sequences of miR-1 mimics and the negative control are presented in Table I . Transfection efficiency was confirmed by analyzing miR-1 expression level using the TaqMan real-time polymerase chain reaction (PCR) system (Applied Biosystems; Thermo Fisher Scientific, Inc., Waltham, MA, USA). Quantitative real-time PCR was run on ABI PRISM 7900HT (Applied Biosystems; Thermo Fisher Scientific, Inc.). Subsequent experimentation was performed 24–48 h after transfection.
7
Vestibular Schwannoma Cell Line Protocol
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The human vestibular schwannoma cell line HEI-193 was obtained from the House Ear Institute (Los Angeles, CA, USA). Cells were cultured in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum at 37°C in a humidified incubator. miR-1 mimics, miR-1 inhibitors (anti-miR-1), and the negative control (NC) were synthesized and purified by GenePharma (Shanghai, China). The VEGFA-expressing vector pcDNA3.1-VEGFA (lacking the VEGFA 3'-UTR) was obtained from Sangon Biotech (Shanghai, China). Transfection experiments were performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA).
8
CRC Cell Lines and miRNA Transfection
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CRC cell lines HT29, HCT116, SW480, and SW620 were purchased from the American Type Culture Collection (ATCC; Manassas, Va) and maintained as previously described [10 (link)]. Additionally, a human CRC cell subline with unique liver metastatic potential, designated SW480/M5, was established in our laboratory [21 (link)] and used in the analysis. The cells were cultured in RPMI 1640 (Hyclone; Logan, Utah, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco-BRL, Invitrogen; Paisley, UK) at a humidity of 5% CO2 at 37°C.
miRNAs were transfected at a working concentration of 100 nmol/L using Lipofectamine 2000 reagent (Invitrogen; Carlsbad, Calif, USA). The miR-1 mimic, a nonspecific miR control, anti-miR-1 (miR-1 inhibitor), and a nonspecific anti-miR control were all purchased from GenePharma (Shanghai, China). Protein and RNA samples were extracted from subconfluent cells during the exponential phase of growth.
miRNAs were transfected at a working concentration of 100 nmol/L using Lipofectamine 2000 reagent (Invitrogen; Carlsbad, Calif, USA). The miR-1 mimic, a nonspecific miR control, anti-miR-1 (miR-1 inhibitor), and a nonspecific anti-miR control were all purchased from GenePharma (Shanghai, China). Protein and RNA samples were extracted from subconfluent cells during the exponential phase of growth.
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