State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
Department of Immunology and Microbiology, School of Life Sciences, Southern University of Science and Technology, Shenzhen 518055, China.
ACS Synth Biol. 2023 Feb 17;12(2):360-368. doi: 10.1021/acssynbio.2c00441. Epub 2023 Jan 20.
To recognize and manipulate a specific microbe of a crowded community is a highly challenging task in synthetic biology. Here we introduce a highly selective protein delivery platform, termed DUEC, which responds to direct contact of attacking cells by engineering the tit-for-tat/dueling response of H1-T6SS (type VI secretion system) in . Using a Cre-recombinase-dependent reporter, we screened H1-T6SS-secreted substrates and developed Tse6 as the most effective secretion tag for Cre delivery. DUEC cells can discriminately deliver the Tse6-Cre cargo into the cytosol of T6SS but not T6SS cells. DUEC could also deliver a nuclease cargo, Tse6-NucSe1, to selectively kill provoking cells in a mixed community. These data demonstrate that the DUEC cell not only is a prototypical physical-contact sensor and delivery platform but also may be coupled with recombination-based circuits with the potential for complex tasks in mixed microbial communities.
在合成生物学中，识别和操纵拥挤群落中的特定微生物是一项极具挑战性的任务。在这里，我们引入了一种高度选择性的蛋白质递送平台，称为 DUEC，它通过工程化 H1-T6SS（类型 VI 分泌系统）的针锋相对/决斗反应来响应攻击细胞的直接接触。使用 Cre 重组酶依赖性报告基因，我们筛选了 H1-T6SS 分泌的底物，并开发了 Tse6 作为 Cre 递送的最有效分泌标签。DUEC 细胞可以将 Tse6-Cre 货物有区别地递送到 T6SS 但不是 T6SS 细胞的细胞质中。DUEC 还可以递送一种核酸酶货物 Tse6-NucSe1，以选择性地杀死混合群落中的挑衅细胞。这些数据表明，DUEC 细胞不仅是一种典型的物理接触传感器和递送平台，而且还可以与基于重组的电路结合使用，从而有可能在混合微生物群落中执行复杂任务。
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