【摘要】 本文开发了一种位于起始密码子附近的发夹元件以在哺乳动物细胞中调节mRNA的表达水平.研究了发夹元件的热力学稳定性、GC含量、与5’帽的距离等参数对mRNA表达的影响.此外,隐藏在发夹元件内的起始密码子和上游开放阅读框也会削弱mRNA的翻译起始.证明了此简单的工程化发夹结构可以有效地用于可控的mRNA翻译调控,有利于在不同类型哺乳动物细胞中实现可预测的基因调控.更多还原

【Abstract】 The hairpin element(HpE) near the start codon in the 5’ untranslated region(UTR) was developed to tune the mRNA translation in mammalian cells.The parameters of HpEs including thermodynamic stability,the content of guanine and cytosine,and distance between HpEs and the5’ cap were investigated.These parameters influenced the mRNA expression level.In addition,the start codon and the upstream open reading frame sequestered within the structures of HpEs also reduced the translation initiation.In summary,this study shows that the simple engineering HpE structure can be efficiently adopted for gene expression regulation.The predictable controllability of this simple cloning strategy can potentially achieve precise gene expression regulation in different mammalian cell types.更多还原