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.本研究旨在探究异源环境下3'非翻译区（3'-UTR）基序的功能。本研究将若干经预测可对信使RNA（mRNA）发挥稳定或去稳定作用的3'非翻译区基序，插入至分别调控报告基因mCherry表达的终止子序列中，所用的表达框架包括：pRps3-mCherry-tRps3、pPgk1-mCherry-tRps3、pTsa1-mCherry-tTsa。本研究采用5'端磷酸化测序（5'P sequencing）技术，旨在明确3'非翻译区的修饰是否会影响报告基因编码区3'末端区域的核糖体动力学过程，以及mRNA的共翻译降解情况。