The rational design and realisation of simple-to-use genetic control elements that are modular, orthogonal and robust is essential to the construction of predictable and reliable biological systems of increasing complexity. To this effect, we introduce modular Artificial RNA interference (mARi), a rational, modular and extensible design framework that enables robust, portable and multiplexed post-transcriptional regulation of gene expression in Escherichia coli. The regulatory function of mARi was characterised in a range of relevant genetic contexts, demonstrating its independence from other genetic control elements and the gene of interest, and providing new insight into the design rules of RNA based regulation in E. coli, while a range of cellular contexts also demonstrated it to be independent of growth-phase and strain type. Importantly, the extensibility and orthogonality of mARi enables the simultaneous post-transcriptional regulation of multi-gene systems as both single-gene cassettes and poly-cistronic operons. To facilitate adoption, mARi was designed to be directly integrated into the modular BASIC DNA assembly framework. We anticipate that mARi-based genetic control within an extensible DNA assembly framework will facilitate metabolic engineering, layered genetic control, and advanced genetic circuit applications.

中文翻译：

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用于多重基因调控的模块化 RNA 干扰系统

模块化、正交和稳健的简单易用的遗传控制元件的合理设计和实现对于构建可预测和可靠的复杂性日益增加的生物系统至关重要。为此，我们引入了模块化人工RNA干扰( mARi )，这是一种合理、模块化和可扩展的设计框架，能够对大肠杆菌中的基因表达进行稳健、便携和多路复用的转录后调控. mARi 的调控功能在一系列相关遗传背景中得到表征，证明其独立于其他遗传控制元件和感兴趣的基因，并为大肠杆菌中基于 RNA 的调控设计规则提供了新的见解，而一系列细胞环境也表明它与生长期和菌株类型无关。重要的是，mARi 的可扩展性和正交性使得多基因系统能够同时作为单基因盒和多顺反子操纵子进行转录后调控。为了促进采用，mARi 被设计为直接集成到模块化 BASIC DNA 组装框架中。我们预计可扩展 DNA 组装框架内基于 mARi 的遗传控制将促进代谢工程、分层遗传控制和先进的遗传电路应用。