microRNAs (miRNAs) are an extensive class of newly identified endogenous small regulatory molecules. Many studies show that miRNAs play a critical role in almost all biological and metabolic progress through targeting protein-coding genes for mRNA cleavage or translation inhibition. Many miRNAs are also identified from cotton using computational and/or experimental approaches, including the next-generation deep sequencing technology. However, their functions are unclear. In this chapter, we describe a detailed method for overexpressing miRNA, miR156 as an example, in cotton using Agrobacterium-mediated genetic transformation. This provides an approach to investigate the function and regulatory mechanism of miRNAs in cotton.
Keywords: Agrobacterium-mediated transformation; Cotton; Gene regulation; Target; microRNA.