Chromosome-free bacterial cells are safe and programmable platforms for synthetic biology
Chromosome-free bacterial cells are safe and programmable platforms for synthetic biology
复制标题
DOI：
10.1073/pnas.1918859117
复制
发表时间：
2020-03-24
影响因子：
11.1
通讯作者：
Huang, Wei E.
中科院分区：
文献类型：
--
作者：
Fan, Catherine;Davison, Paul A.;Huang, Wei E.
一种称为SimCells（简单细胞）的无染色体细胞已经从大肠杆菌、恶臭假单胞菌和真养罗尔斯通氏菌中产生。这些细菌的天然染色体的去除是通过异源I-CeuI核酸内切酶和内源性核酸酶的降解活性产生的双链断裂来实现的。我们已经证明，细胞机器在这些无染色体的SimCells中保持功能，并且能够处理各种遗传电路。这包括糖酵解途径（由10个基因组成）和可诱导的遗传回路。发现糖酵解途径由于其再生ATP和NADH/NADPH的能力而显著延长了SimCells的寿命。SimCells能够在染色体去除后连续表达合成的遗传电路10 d。作为原理证明，我们证明了SimCells可以用作细菌治疗的安全试剂（因为它们不能复制）。SimCells用于从水杨酸合成儿茶酚（一种有效的抗癌药物），以抑制肺、脑和软组织癌细胞。SimCells代表了一种简化的合成生物学底盘，可以编程以安全地制造和交付产品，而不会受到宿主基因组的干扰。
A type of chromosome-free cell called SimCells (simple cells) has been generated from Escherichia coli, Pseudomonas putida, and Ralstonia eutropha. The removal of the native chromosomes of these bacteria was achieved by double-stranded breaks made by heterologous I-CeuI endonuclease and the degradation activity of endogenous nucleases. We have shown that the cellular machinery remained functional in these chromosome-free SimCells and was able to process various genetic circuits. This includes the glycolysis pathway (composed of 10 genes) and inducible genetic circuits. It was found that the glycolysis pathway significantly extended longevity of SimCells due to its ability to regenerate ATP and NADH/NADPH. The SimCells were able to continuously express synthetic genetic circuits for 10 d after chromosome removal. As a proof of principle, we demonstrated that SimCells can be used as a safe agent (as they cannot replicate) for bacterial therapy. SimCells were used to synthesize catechol (a potent anticancer drug) from salicylic acid to inhibit lung, brain, and soft-tissue cancer cells. SimCells represent a simplified synthetic biology chassis that can be programmed to manufacture and deliver products safely without interference from the host genome.