The 3′-UTR sequence of PVT1 was amplified from normal human genomic DNA and subcloned into the pRL-CMV luciferase reporter vector (Ambion, Austin, TX, USA). HEK293T cells were seeded at a density of 5 × 103 cells/well in 96-well plates and cotransfected with firefly luciferase target reporter (50 ng) and pRL-CMV Renilla luciferase control reporter (5 ng) and miRNA mimics or negative control using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions. After incubation for 48 h, the firefly and Renilla luciferase vitality were assayed using the Dual Luciferase System (E1910; Promega, Madison, WI, USA).
Prl cmv luciferase reporter vector
Manufactured by Thermo Fisher Scientific
Sourced in United States
The PRL-CMV luciferase reporter vector is a plasmid construct designed for the expression and detection of luciferase reporter activity in mammalian cell lines. The vector contains the cytomegalovirus (CMV) promoter to drive the expression of the luciferase gene, which can be used as a reporter for gene expression or transcriptional activity studies.
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Lab products found in correlation
2 protocols using prl cmv luciferase reporter vector
1
Luciferase Reporter Assay for PVT1 3'-UTR
2
Characterizing NEAT1 and KLF3 3'-UTR Regulation
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We amplified the 3′-UTR sequence of NEAT1 and KLF3 in normal human genomic DNA. Then, we subcloned the cells in pRL-CMV luciferase reporter vector (Ambion, USA). We seeded HEK293T cells at a density of 5 × 103 cells in each well of the 96-well plates. Then, we co-transfected the cells with firefly luciferase target reporter and pRL-CMV Renilla luciferase control reporter. We co-transfected the miRNA mimics group or negative control group using Lipofectamine 2000 (Invitrogen, USA). We incubated the assay for 48 h and evaluated the luciferase activity through the Dual-Luciferase System (Promega, USA).
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