The advances in synthetic biology bring exciting new opportunities to reprogram microorganisms with novel functionalities for environmental applications. For real‐world applications, a genetic tool that enables genetic engineering in a stably genomic inherited manner is greatly desired. In this work, we design a novel genetic device for rapid and efficient genome engineering based on the intron‐encoded homing‐endonuclease empowered genome editing (iEditing). The iEditing device enables rapid and efficient genome engineering in Shewanella oneidensis MR‐1, the representative strain of the electroactive bacteria group. Moreover, combining with the Red or RecET recombination system, the genome‐editing efficiency was greatly improved, up to approximately 100%. Significantly, the iEditing device itself is eliminated simultaneously when genome editing occurs, thereby requiring no follow‐up to remove the encoding system. Then, we develop a new extracellular electron transfer (EET) engineering strategy by programming the parallel EET systems to enhance versatile EET. The engineered strains exhibit sufficiently enhanced electron output and pollutant reduction ability. Furthermore, this device has demonstrated its great potential to be extended for genome editing in other important microbes. This work provides a useful and efficient tool for the rapid generation of synthetic microorganisms for various environmental applications.

中文翻译：

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使用新型iEditing设备进行快速高效的基因组工程设计，可对电活性细菌的多种细胞外电子转移进行编程

合成生物学的进步带来了令人兴奋的新机会，可以对具有新型功能的微生物进行重编程以用于环境应用。对于现实世界的应用，非常需要一种能够以稳定的基因组遗传方式进行基因工程的遗传工具。在这项工作中，我们设计基于所述用于快速和有效的基因组工程的新的遗传元件我ntron编码归巢内切核酸酶的基因组增效编辑（iEditing）。iEditing设备可实现快速高效地对印度希瓦氏菌进行基因组工程MR-1，代表电活性细菌群。此外，结合Red或RecET重组系统，基因组编辑效率得到了极大的提高，达到了大约100％。重要的是，在进行基因组编辑时，可以同时消除iEditing设备本身，因此无需进行后续操作即可删除编码系统。然后，我们通过对并行EET系统进行编程以增强通用EET，从而开发了一种新的细胞外电子转移（EET）工程策略。工程菌株表现出充分增强的电子输出和污染物减少能力。此外，该设备已展示出其巨大潜力，可以扩展到其他重要微生物的基因组编辑中。