5PSeq supporting study Probing 3'UTRs as modular regulators of gene expression

The goal of the study is to analyse the function of 3'-UTR motifs in the non-native context. Several 3'-UTR motifs predicted to have stabilizing or destabilizing effects on mRNA were inserted into terminators controlling the expression of reporter mCherry gene in following contexts: pRps3-mCherry-tRps3, pPgk1-mCherry-tRps3, pTsa1-mCherry-tTsa. 5'P sequencing was used to determine whether or not modifications in the 3'-UTR region affect ribosomal dynamics and co-translational degradation of mRNA in the 3' end region of reporter coding sequence. Overall design: 5'P sequencing of Saccharomyces cerevisiae strains expressing mCherry reporter controlled by modified and non-modified terminators. mCherry reporter constructs are expressed from a low copy number plasmid. All modified constructs have three 9bp insertion sites with different combinations of the following motifs: N, a randomly generated sequence; A, an ATATTC motif; H, a TTTCATTTC motif; and T, a TGTACAATA motif.