Cells were transfected with 1:1 ratio of plasmids encoding firefly luciferase with varied poly(A) sequence lengths within the 5′ UTR and a control Renilla luciferase plasmid (Addgene) using Lipofectamine 2000. Luciferase activity was measured using a Dual-Glo reagent kit (Promega) and a GloMax (Promega) luminometer, following the manufacturer’s instructions.
Renilla luciferase plasmid
Manufactured by Addgene
The Renilla luciferase plasmid is a laboratory tool used to measure gene expression. It contains the Renilla luciferase gene, which encodes an enzyme that produces bioluminescence when exposed to a substrate. This plasmid is commonly used as a reporter for monitoring promoter activity or studying gene regulation in cell-based assays.
Automatically generated - may contain errors
2 protocols using renilla luciferase plasmid
1
Luciferase Assay with Varied 5' UTR
2
Transcriptional Regulation via YAP1-TEAD2 Axis
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A 500 bp region spanning the Jdp2 or control enhancers (Supplementary Information) were cloned from Hepa1-6 genomic DNA into a firefly luciferase reporter (pBV-luc; Addgene #16539), followed by the SV40 promoter. TEAD2 plasmid was generated by subcloning the TEAD2 sequence from Addgene #33107 into lentiCas9-blast (Addgene #52962) to replace the Cas9 sequence. 293fs cells were plated (6-well) and transfected with 800 ng of YAP1 plasmid (Addgene #86497), 800 ng of TEAD2 plasmid, 400 ng of firefly luciferase reporter plasmid, and 400 ng of Renilla luciferase plasmid (Addgene #27163) using lipofectamine 3000 (Invitrogen) for 72 hours. Luciferase activity was measured using the Dual-glo luciferase assay system (Promega) according to the manufacturer's protocol. Finally, we normalized firefly luciferase activity to Renilla.
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