The coding sequences of CD276 were cloned into pcDNA3.1 (+) to generate CD276 expression vectors. The wild-type CD276 3′UTR was cloned into the pMIR-REPORT luciferase vector (Ambion, Austin, TX, USA). Mutant CD276 3′UTR was generated based on the pMIR-CD276-3′UTR by mutating 3 nt that are recognized by miR-187. The primers for CD276 were: 5′-TGTGGATCCCTGTCATCTGGGAAGTAACAACGCA-3′ (forward) and 5′-AAGTCTAGAGAGCCACTACTGCCTGTTGTCTTTG-3′ (reverse). The primers for CD276 3′UTR were: 5′-TCTGAGCTCGCTAAACAGCCATAAACGGAAACGC-3′ (forward) and 5′-ACCACGCGTGCGTAGATTCTCCTTTATGGGGCTG-3′ (reverse). MiR-187 mimics, miR mimic control, miR-187 inhibitor, miR inhibitor control, and small interfering RNA targeting CD276 were purchased from GenePharma (Shanghai, China). Transfection was performed according to the manufacturer's instructions.
Small interfering rna targeting
Manufactured by GenePharma
Sourced in China
Small interfering RNA (siRNA) targeting is a laboratory technique used to silence or downregulate the expression of specific genes. siRNA molecules are designed to complementarily bind to target mRNA, leading to its degradation or translational repression, thereby reducing the production of the corresponding protein.
Automatically generated - may contain errors
Lab products found in correlation
Anti mir nc, by RiboBio (3 mentions)
Si xist, by GenePharma (3 mentions)
Anti mir 497 5p, by RiboBio (3 mentions)
Pcdna3.1 vector vector, by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (3 mentions)
Si nc, by GenePharma (3 mentions)
Mir nc, by RiboBio (3 mentions)
Pmir report luciferase vector, by Thermo Fisher Scientific (1 mentions)
Mir mimic control, by GenePharma (1 mentions)
Mir 187 mimics, by GenePharma (1 mentions)
Mir 187 inhibitor, by GenePharma (1 mentions)
Mir inhibitor control, by GenePharma (1 mentions)
4 protocols using small interfering rna targeting
1
CD276 Expression Regulation by miR-187
2
Regulation of XIST and miR-497-5p in CRC
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Four CRC cell lines (including LOVO, SW480, HT-29, and HCT-116) and one normal colon epithelial cell line (FHC) were obtained from Shanghai Institute of Biochemistry and Cell Biology (Shanghai, China). All cell lines were cultured in DMEM (Gibco, USA) supplemented with 10% fetal bovine serum (FBS; Tianhang, Hangzhou, China) and 1% penicillin-streptomycin in a humidified incubator (37 °C, 5% CO2). Small interfering RNA targeting XIST (si-XIST: 5′-GCCCUUCUCUUCGAACUGUTT-3′) and its matching control (si-NC: 5′-CGTTAATCGCGTATAATACGCGTAT-3′), were obtained from Genepharma (Shanghai, China). A miR-497-5p mimic (miR-497-5p), its inhibitor (anti-miR-497-5p), and their corresponding controls (miR-NC and anti-miR-NC) were obtained from Ribobio (Guangzhou, China). For overexpression vectors, the sequence of FOXK1 was cloned into the pcDNA3.1 vector (vector) according to the manufacturer’s instructions (Life Technologies). Briefly, 100 nM of miR-497-5p mimics or miR-497-5p inhibitor and 1000 ng plasmid were transfected into each 6-well plate for 48 h using Lipofectamine 3000 (Invitrogen, Waltham, MA, USA). For animal experiments, cells were stably transfected with an XIST lentiviral vector, constructed by Hanyin Biotechnology Co., Ltd., Shanghai, China. After transfection, DMEM medium was replaced with DMEM supplemented with puromycin (3 µg/mL) as a positive selection for infected cells.
3
XIST and miR-497-5p Regulation in Colorectal Cancer
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Four CRC cell lines (including LOVO, SW480, HT-29, and HCT-116) and one normal colon epithelial cell line (FHC) were obtained from Shanghai Institute of Biochemistry and Cell Biology (Shanghai, China). All cell lines were cultured in DMEM (Gibco, USA) supplemented with 10% fetal bovine serum (FBS; Tianhang, Hangzhou, China) and 1% penicillin-streptomycin in a humidi ed incubator (37°C, 5% CO 2 ). Small interfering RNA targeting XIST (si-XIST: 5'-GCCCUUCUCUUCGAACUGUTT-3') and its matching control (si-NC: 5'-CGTTAATCGCGTATAATACGCGTAT-3'), were obtained from Genepharma (Shanghai, China). A miR-497-5p mimic (miR-497-5p), its inhibitor (anti-miR-497-5p), and their corresponding controls (miR-NC and anti-miR-NC) were obtained from Ribobio (Guangzhou, China). For overexpression vectors, the sequence of FOXK1 was cloned into the pcDNA3.1 vector (vector) according to the manufacturer's instructions (Life Technologies). Brie y, 100 nM of miR-497-5p mimics or miR-497-5p inhibitor and 1000 ng plasmid were transfected into each 6-well plate for 48 h using Lipofectamine 3000 (Invitrogen, Waltham, MA, USA). For animal experiments, cells were stably transfected with an XIST lentiviral vector, constructed by Hanyin Biotechnology Co., Ltd., Shanghai, China. After transfection, DMEM medium was replaced with DMEM supplemented with puromycin (3 µg/mL) as a positive selection for infected cells.
4
XIST and miR-497-5p Regulation in Colorectal Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Four CRC cell lines (including LOVO, SW480, HT-29, and HCT-116) and one normal colon epithelial cell line (FHC) were obtained from Shanghai Institute of Biochemistry and Cell Biology (Shanghai, China). All cell lines were cultured in DMEM (Gibco, USA) supplemented with 10% fetal bovine serum (FBS; Tianhang, Hangzhou, China) and 1% penicillin-streptomycin in a humidi ed incubator (37°C, 5% CO 2 ). Small interfering RNA targeting XIST (si-XIST: 5'-GCCCUUCUCUUCGAACUGUTT-3') and its matching control (si-NC: 5'-CGTTAATCGCGTATAATACGCGTAT-3'), were obtained from Genepharma (Shanghai, China). A miR-497-5p mimic (miR-497-5p), its inhibitor (anti-miR-497-5p), and their corresponding controls (miR-NC and anti-miR-NC) were obtained from Ribobio (Guangzhou, China). For overexpression vectors, the sequence of FOXK1 was cloned into the pcDNA3.1 vector (vector) according to the manufacturer's instructions (Life Technologies). Brie y, 100 nM of miR-497-5p mimics or miR-497-5p inhibitor and 1000 ng plasmid were transfected into each 6-well plate for 48 h using Lipofectamine 3000 (Invitrogen, Waltham, MA, USA). For animal experiments, cells were stably transfected with an XIST lentiviral vector, constructed by Hanyin Biotechnology Co., Ltd., Shanghai, China. After transfection, DMEM medium was replaced with DMEM supplemented with puromycin (3 µg/mL) as a positive selection for infected cells.
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