Analyzing the mechanism of stable mRNA 3’-UTR sequences

Abstract

mRNAs are widely studied for applications of vaccines and therapeutics. However, the instability of mRNAs is still the major challenge for the development of mRNA as a therapeutic agent. The stability of an mRNA is dependent on the nucleotide sequences and secondary structures. Particularly, 3′-untranslated region (3′-UTR) is well known as the regulatory element of mRNA stability. Previous studies have discovered some stable 3′-UTR sequences and structures. However, most of the studies have not analyzed the mechanism underlying the stability regulation of such UTRs. In this study, the stability regulatory mechanism of the known stable 3′-UTR sequences is analyzed with respect to the RNA binding proteins. The stability of mRNAs is regulated post-tranascriptionally through the interaction with the RNA binding proteins. Recently, some RNA binding proteins that stabilize mRNAs and enhance translation have been discovered but not well studied yet. Through 3′ end biotinylation of known stable RNA sequences, in vitro RNA-RBP binding assay is performed. To further investigate the interactome of stable 3′-UTRs in cells, MBSV6 and MBSV6-coat protein system is used in HEK-293T cells.