International Centre for Genetic Engineering and Biotechnology, Padriciano 99, 34149 Trieste, Italy.
Mol Biotechnol. 2011 Jul;48(3):290-7. doi: 10.1007/s12033-011-9390-x.
Eukaryotic gene expression relies on several complex molecular machineries that act in a highly coordinated fashion. These machineries govern all the different steps of mRNA maturation, from gene transcription and pre-mRNA processing in the nucleus to the export of the mRNA to the cytoplasm and its translation. In particular, the pre-mRNA splicing process consists in the joining together of sequences (known as "exons") that have to be differentiated from their intervening sequences commonly referred to as "introns." The complex required to perform this process is a very dynamic macromolecular ribonucleoprotein assembly that functions as an enzyme, and is called the "spliceosome." Because of its flexibility, the splicing process represents one of the main mechanisms of qualitative and quantitative regulation of gene expression in eukaryotic genomes. This flexibility is mainly due to the possibility of alternatively recognizing the various exons that are present in a pre-mRNA molecule and therefore enabling the possibility of obtaining multiple transcripts from the same gene. However, regulation of gene expression by the spliceosome is also achieved through its ability to influence many other gene expression steps that include transcription, mRNA export, mRNA stability, and even protein translation. Therefore, from a biotechnological point of view the splicing process can be exploited to improve production strategies and processes of molecules of interest. In this work, we have aimed to provide an overview on how biotechnology applications may benefit from the introduction of introns within a sequence of interest.
真核基因表达依赖于几种复杂的分子机制,这些机制以高度协调的方式发挥作用。这些机制控制着 mRNA 成熟的所有不同步骤,从细胞核中的基因转录和前体 mRNA 处理到 mRNA 向细胞质的输出及其翻译。特别是,前体 mRNA 的剪接过程包括将序列(称为“外显子”)连接在一起,这些序列必须与它们之间的序列区分开来,通常称为“内含子”。执行此过程所需的复杂的是一种非常动态的大分子核糖核蛋白组装体,作为一种酶起作用,称为“剪接体”。由于其灵活性,剪接过程是真核基因组中基因表达定性和定量调控的主要机制之一。这种灵活性主要归因于能够以不同的方式识别存在于前体 mRNA 分子中的各种外显子,从而能够从同一个基因获得多个转录本。然而,剪接体对基因表达的调控也是通过其影响许多其他基因表达步骤的能力来实现的,包括转录、mRNA 输出、mRNA 稳定性,甚至蛋白质翻译。因此,从生物技术的角度来看,可以利用剪接过程来改进感兴趣分子的生产策略和过程。在这项工作中,我们旨在概述生物技术应用如何受益于在感兴趣的序列中引入内含子。