A pmiR-RB-REPORT™ vector (Ribobio Co., Guangzhou, China) was used for 3′ UTR-luciferase reporter assays to detect interactions of miR-663a with JunD (Gene ID: 3727). The TargetScan Human database 6.2 http://targetscan.org/was used to identify miRNA binding sites. The Wild-type miR-663a target site in JunD 3′-untranslated region was CCCCGCC. The mutant miR-663a target site was CGGGCG C. The miR-663a mimic, miR-663a inhibitor and their corresponding negative controls (Ribobio Co. Guangzhou, China) were transfected with pmiR-RB-REPORT™-target gene UTR. Reporter assays were conducted in triplicate. The cells were co-transfected with 100 nM mimic and 100 ng/mL reporter plasmid by attractene agent (Qiagen, Hilden, Germany) according to the manufacturer’s instructions
Mir 663a mimic
Manufactured by RiboBio
Sourced in China
The MiR-663a mimic is a synthetic RNA molecule that mimics the sequence and function of the naturally occurring microRNA MiR-663a. MicroRNAs are small, non-coding RNA molecules that play a role in the regulation of gene expression. The MiR-663a mimic can be used in research applications to study the biological functions and effects of this specific microRNA.
Automatically generated - may contain errors
Lab products found in correlation
Mir 663a inhibitor, by RiboBio (3 mentions)
Pmir rb report vector, by RiboBio (1 mentions)
Pmir rb report, by RiboBio (1 mentions)
Bxpc 3, by American Type Culture Collection (1 mentions)
Inhibitor control, by RiboBio (1 mentions)
Control sirna, by RiboBio (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Mir nc, by RiboBio (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Panc 1, by American Type Culture Collection (1 mentions)
Aspc 1, by American Type Culture Collection (1 mentions)
Sw1990, by American Type Culture Collection (1 mentions)
Paca 2, by American Type Culture Collection (1 mentions)
Hpde6 c7, by American Type Culture Collection (1 mentions)
Pmir glo vector, by Tsingke (1 mentions)
Pla2g6, by Tsingke (1 mentions)
3 protocols using mir 663a mimic
1
miR-663a Regulates JunD Expression via 3'-UTR Binding
2
Pancreatic Cancer Cell Line Characterization
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The human pancreatic cancer cell lines AsPC-1, BxPC-3, PANC-1, PaCa-2, and SW1990, and the human pancreatic ductal epithelial cell line HPDE6-C7 were from ATCC (Rockville, MD, United States). The cells were cultured in RPMI-1640 or DMEM (Gibco, Waltham, MA, United States) containing 10% fetal bovine serum (FBS). Overexpression plasmid containing C9orf139 (sh-C9orf139), plasmids for knockdown (si-C9orf139-#1, si-C9orf139-#2, and si-C9orf139-#3), and Sox12 expression plasmid (pcDNA3.1-Sox12) were constructed by Shanghai GenePharma Co., Ltd. Sox12 shRNA, Sox12 siRNA, control siRNA, miR-663a mimic, miR-NC, miR-663a inhibitor, and control inhibitor were obtained from RiboBio (Guangzhou, China). Cell transfection was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, United States) according to the manufacturer’s instructions.
3
Identifying miR-663a Regulation of PLA2G6
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MiR-663a-mimic, miR-663a-inhibitor and the corresponding negative control were synthesized by RiboBio. The double-stranded siRNA and siNC were synthesized and high performance purified (GenePharma). The targeted and negative control siRNA for PLA2G6 mRNA were as follows: PLA2G6: 5′-GCCUAAACAACCUAGAACUTT-3′ (sense); Control: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense). The wild type (WT) and mutated (MUT) 3′-UTR sequences of PLA2G6 were purchased from Tsingke (Nanjing, China), which were predicted to interact with miR-663a, were cloned into a firefly luciferase-expressing pmiR-GLO vector (Tsingke) to acquire the PLA2G6 3′-UTR reporter constructs (pmiR-PLA2G6-WT and pmiR-PLA2G6-MUT).
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