Pseudomonas putida is a highly attractive production system for industrial needs. However, for its improvement as a biocatalyst at the industrial level, modulation of its gene expression is urgently needed. We report the construction of a plasmid expressing a small RNA-based system with the potential to be used for different purposes. Due to the small RNAs modular composition, the design facilities and ability to tune gene expression, they constitute a powerful tool in genetic and metabolic engineering. In the tool presented here, customized sRNAs are expressed from a plasmid and specifically directed to any region of a chosen target. Expression of these customized sRNAs is shown to differentially modulate the level of endogenous and heterologous reporter genes. The antisense interaction of the sRNA with the mRNA produces different outcomes. Depending on the particularity of each sRNA-target mRNA pair, we demonstrate the duality of this system, which is able either to decrease or increase the expression of the same given gene. This system combines high specificity with the potential to be widely applied, due to its predicted ability to modulate the expression of virtually any given gene. This plasmid can be used to redesign P. putida metabolism, fulfilling an important industrial gap.

中文翻译：

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量身定制的sRNA：一种质粒工具，用于控制恶臭假单胞菌中目标mRNA的表达。

恶臭假单胞菌（Pseudomonas putida）是满足工业需求的极具吸引力的生产系统。然而，为了在工业水平上改进其作为生物催化剂，迫切需要调节其基因表达。我们报告了一种表达基于小RNA的系统的质粒的构建，其具有用于不同目的的潜力。由于小分子RNAs的组成，设计设施和调节基因表达的能力，它们构成了基因和代谢工程的有力工具。在此处介绍的工具中，定制的sRNA从质粒表达，并特异性针对所选靶标的任何区域。这些定制的sRNA的表达显示出差异地调节内源和异源报道基因的水平。sRNA与mRNA的反义相互作用产生不同的结果。取决于每个sRNA靶mRNA对的特殊性，我们证明了该系统的双重性，它能够减少或增加相同给定基因的表达。由于该系统具有调节几乎任何给定基因表达的预测能力，因此该系统将高特异性与广泛应用的潜力相结合。该质粒可用于重新设计恶臭假单胞菌的代谢，从而弥补重要的工业差距。