Chromosome-free bacterial cells are safe and programmable platforms for synthetic biology.

Proceedings of the National Academy of Sciences of the United States of America
Catherine FanWei E Huang

Abstract

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 fr...Continue Reading

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Citations

Jul 10, 2020·Journal of Industrial Microbiology & Biotechnology·Zhu LiDawei Zhang
May 8, 2021·Microbial Biotechnology·Xinyi WanBaojun Wang
Sep 6, 2020·New Biotechnology·Víctor de LorenzoMarkus Schmidt

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Methods Mentioned

BETA
flow cytometry
Fluorescence
biosensors
fluorescence microscopy
Assay

Software Mentioned

SimCell
SimCells
MassIVE

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