Rapid Cloning and Validation of MicroRNA Shuttle Vectors: A Practical Guide

MicroRNAs (miRNAs) have emerged as important modulators of eukaryotic gene expression through a process called RNA interference (RNAi). Over the last several years, a large amount of work has focused on understanding how miRNAs are expressed and processed to a biologically functional form. This knowledge has enabled the development of RNAi as a molecular tool for investigating basic biological questions or as a therapeutic technique. Artificial miRNA shuttle vectors can be engineered to mimic natural miRNAs and subsequently used to suppress any gene of interest. Here, we describe a simple method to build and functionally validate artificial miRNA shuttles.