荧光素酶报告基因检测法测定干扰素的信号转导活性。

Luciferase Reporter Assay for Determining the Signaling Activity of Interferons.

机构信息

Institutes of Biology and Medical Sciences, Soochow University, Suzhou, Jiangsu, China.

Institute/Department of Laboratory Medicine, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, China.

出版信息

Methods Mol Biol. 2025;2854:19-28. doi: 10.1007/978-1-0716-4108-8_3.

DOI:10.1007/978-1-0716-4108-8_3

PMID:39192114

Abstract

The classic dual luciferase reporter assay has been widely used to rapidly and accurately determine the transcriptional activity of a given promoter induced by certain signal pathways in the cells. In particular, the sensitive characteristics of luciferase highlight its significance in many experiments, such as weak promoter analysis, transfection studies using small amounts of DNA, and detection in cell lines with low transfection efficiency. This chapter presents detailed information and experimental procedures for measuring interferon (IFN)-induced Interferon-Stimulated Response Element (ISRE) promoter activity using the dual luciferase reporter assay.

摘要

经典的双荧光素酶报告基因检测实验已被广泛用于快速、准确地测定细胞内特定信号通路诱导的特定启动子的转录活性。特别是，荧光素酶的高灵敏度特点使其在许多实验中具有重要意义，如弱启动子分析、使用少量 DNA 的转染研究，以及低转染效率细胞系的检测。本章提供了使用双荧光素酶报告基因检测实验测量干扰素（IFN）诱导的干扰素刺激反应元件（ISRE）启动子活性的详细信息和实验步骤。