Department of Molecular, Cell and Systems Biology, Institute for Integrative Genome Biology, University of California, Riverside, CA, USA.
Methods Mol Biol. 2021;2170:101-115. doi: 10.1007/978-1-0716-0743-5_7.
The luciferase reporter assay is a widely used tool to study the cis and trans factors controlling regulation of gene expression. In this assay, regulatory elements can be fused to the luciferase gene, and as a result effect protein output by changing rates of transcription, rates of translation, or mRNA stability. This protocol focuses on probing the function of RNA-binding proteins (RBPs) through their interactions with the 3' untranslated region (UTR), thus examining gene expression regulation on the mRNA level. Assessment of 3' UTR sequence requirements, as well as single and co-regulatory roles of RBPs in regulation of mRNAs will be discussed.
荧光素酶报告基因检测是一种广泛用于研究顺式和反式因子调控基因表达的工具。在该检测中，调控元件可以与荧光素酶基因融合，从而通过改变转录、翻译或 mRNA 稳定性的速率来影响蛋白的输出。本方案主要关注通过与 3'非翻译区 (UTR) 的相互作用来研究 RNA 结合蛋白 (RBP) 的功能，从而在 mRNA 水平上检测基因表达调控。本文将讨论 3'UTR 序列要求，以及 RBP 在调节 mRNA 中的单一和协同调控作用。
