Luciferase Assay for miR-31-5p Regulation

The wild type or mutant 3′-UTR of Cab39 was cloned into the pGL3 plasmid (Promega; Madison, WI, USA) containing a luciferase report gene, which was then cotransfected with the miR-31-5p agomir or AgNC using the Lipofectamine 6000 Reagent (Beyotime; Shanghai, China). The cells were cultured for 48 h and then collected to detect the firefly and Renilla luciferase activities by a Dual-Luciferase Reporter Assay System (Promega; Madison, WI, USA) [31 (link)–33 (link)].

Free full text: Click here

Jiang W.L., Zhao K.C., Yuan W., Zhou F., Song H.Y., Liu G.L., Huang J., Zou J.J., Zhao B, & Xie S.P. (2020). MicroRNA-31-5p Exacerbates Lipopolysaccharide-Induced Acute Lung Injury via Inactivating Cab39/AMPKα Pathway. Oxidative Medicine and Cellular Longevity, 2020, 8822361.

Publication 2020

pGL3 plasmid Lipofectamine 6000 Reagent Dual-Luciferase Reporter Assay System

Assay Cells Firefly Gene Lipofectamine Luciferase Pgl3 Plasmid Promega Renilla luciferase

Corresponding Organization : Renmin Hospital of Wuhan University

Other organizations : Huazhong University of Science and Technology

Top 5 similar protocols

Variable analysis

independent variables

Wild type or mutant 3'-UTR of Cab39 cloned into the pGL3 plasmid
MiR-31-5p agomir or AgNC

dependent variables

Firefly and Renilla luciferase activities

control variables

Cells were cultured for 48 h

controls

Positive control: Wild type 3'-UTR of Cab39 cloned into the pGL3 plasmid
Negative control: AgNC (negative control for miR-31-5p agomir)

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!