First, the luciferase reporter plasmids Nampt-WT and Nampt-Mut was constructed, then cells were co-transfected with above plasmids and NC mimic or miR-377-3p mimic respectively by using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA). After co-incubation for 48h, cells were collected for lysis and luciferase activity was measured by the dual luciferase reporting and detection system (Promega, WI, USA). Similarly, the protocol also used to explore the combined effects of NEAT1 and miR-377-3p.
Dual luciferase reporting and detection system
Manufactured by Promega
Sourced in United States
The Dual Luciferase Reporting and Detection System is a laboratory tool used for the quantitative measurement of gene expression. It utilizes two different luciferase reporter enzymes to provide a reliable and sensitive method for analyzing transcriptional activity in cells. The system enables simultaneous measurement of two reporter activities within a single sample, which can be used to normalize experimental data.
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Lab products found in correlation
3 protocols using dual luciferase reporting and detection system
1
Investigating miR-377-3p Regulation of Nampt
2
Dual-Luciferase Reporter Assay for ATG12 and AMBRA1
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The pmirGLO dual-luciferase plasmid with wild-type (WT) or mutated (MUT) ATG12 and AMBRA1 3′ UTRs inserted downstream of the firefly luciferase gene was constructed by Generay (Shanghai, China). The luciferase reporter assay was performed according to the instructions of the Dual Luciferase Reporting and Detection System (Promega, Madison, WI, USA).
3
Dual-Luciferase Assay for miRNA-3'UTR Interactions
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HEK-293 T cells were transfected with MEK4–3′UTR-WT, MEK4–3′UTR-MU, JNK1–3′UTR-WT and JNK1–3′UTR-MU plasmids. At the same time, the four groups of cells were transfected with either NC mimic or miR-25-3p mimic to form eight experimental groups. Finally, each group of cells was transfected with the Renilla luciferase plasmid. After 36 h, the cells were harvested for luciferase analysis using a dual luciferase reporting and detection system (Promega, USA). The results were normalized against Renilla luciferase activity.
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