Cell-free system that recapitulates miRNA-mediated mRNA regulation.
Regulation of translation and mRNA degradation are based on a complex interplay of canonical factors, diverse RNA-binding proteins, and non-coding regulatory RNAs (e.g. microRNAs). We have established an in vitro experimental system to recapitulate mechanisms of miRNA-mediated post-transcriptional control by utilizing a cell-free translation technique.
In this system, by adding hairpin-like miRNA precursors or siRNA type double-stranded RNAs, a guide strand of miRNAs are incorporated into Argonaute proteins, and miRISC complexes are formed. The miRISCs bind to the target mRNAs and induce deadenylation and translation inhibition that eventually leads repression of protein synthesis. Our cell-free system is a powerful tool in elucidating miRNA mechanisms.