MicroRNA-mediated deadenylation in a mammalian cell- free system

Abstract

MicroRNAs (miRNAs) are 21–22 nucleotide small noncoding RNAs that regulate gene expression posttranscriptionally. The miRNA is incorporated into the miRNP effector complex. The miRNP complex binds to the mRNA containing the target sites, which are partially homologous to the miRNA sequence, and represses protein synthesis. One of the critical functions of miRNP is the recruitment of deadenylase complexes to the target mRNAs. Deadenylation causes translational inhibition as well as mRNA destabilization. In this chapter, we describe our method to recapitulate miRNA-mediated deadenylation in a mammalian cell-free system.