Ko Hae Young, Lee Young Sik, Kim Soonhag
Laboratory of Molecular Imaging, Department of Biomedical Science, College of Life Science, CHA University, Seoul, Republic of Korea.
Methods Mol Biol. 2014;1098:85-95. doi: 10.1007/978-1-62703-718-1_7.
MicroRNAs (miRNAs) are an abundant class of small noncoding RNA molecules that inhibit the expression of cognate genes in multicellular organisms. These small RNAs have been demonstrated to play crucial roles in a variety of biological processes including cell differentiation, proliferation, and survival. Knowledge of specific expression patterns of miRNAs is critical for functional studies. Here, we describe a bioluminescence reporter gene-based method to measure miRNA activity in cultured cells and mice using a Gaussia luciferase reporter gene controlled by miRNA binding sites in its 3'untranslated region. This method can be used to noninvasively monitor the expression patterns of functionally active miRNAs involved in different biological processes or diseases in mice.
微小RNA（miRNA）是一类丰富的小型非编码RNA分子，可抑制多细胞生物中同源基因的表达。这些小RNA已被证明在多种生物学过程中发挥关键作用，包括细胞分化、增殖和存活。了解miRNA的特定表达模式对于功能研究至关重要。在这里，我们描述了一种基于生物发光报告基因的方法，该方法使用位于其3'非翻译区的miRNA结合位点控制的高斯荧光素酶报告基因来测量培养细胞和小鼠中的miRNA活性。该方法可用于无创监测参与小鼠不同生物学过程或疾病的功能活性miRNA的表达模式。
