Construction and phenotypic classification of synthetic dual-pole Escherichia coli cells

Abstract Macromolecular complexes anchored at one cell pole have previously been utilized to achieve synthetic asymmetric cell division in Escherichia coli. In nature, however, robust asymmetric cell division often relies on gradients established between two opposing poles. Here, we present a second...

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Main Authors: Tsung-Wei Chou, Ing-Jen Huang, Ya-Chu Hsu, Szu-Hua Wu, Jui-Chung Hong, Yu-Hao Ko, Ting-Ran Su, Jie-Hong Roland Jiang, Hsiao-Chun Huang
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Communications Biology
Online Access:https://doi.org/10.1038/s42003-025-08495-w
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author Tsung-Wei Chou
Ing-Jen Huang
Ya-Chu Hsu
Szu-Hua Wu
Jui-Chung Hong
Yu-Hao Ko
Ting-Ran Su
Jie-Hong Roland Jiang
Hsiao-Chun Huang
author_facet Tsung-Wei Chou
Ing-Jen Huang
Ya-Chu Hsu
Szu-Hua Wu
Jui-Chung Hong
Yu-Hao Ko
Ting-Ran Su
Jie-Hong Roland Jiang
Hsiao-Chun Huang
author_sort Tsung-Wei Chou
collection DOAJ
description Abstract Macromolecular complexes anchored at one cell pole have previously been utilized to achieve synthetic asymmetric cell division in Escherichia coli. In nature, however, robust asymmetric cell division often relies on gradients established between two opposing poles. Here, we present a second-generation synthetic genetic circuit that enables two distinct scaffolding proteins to localize at opposite poles in E. coli. By engineering a PodJ fragment as the second scaffold and integrating SpmX, a negative regulator of PodJ condensation, as a direct chimera with PopZ, we effectively confined PopZ and PodJ to opposite poles within E. coli. Additionally, we developed a neural network-based classification method to identify these synthetic dual-pole cells, further advancing the synthetic biology toolkit for programmable cell differentiation.
format Article
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institution Kabale University
issn 2399-3642
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publisher Nature Portfolio
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spelling doaj-art-563b1ce1dc944a209d705fb576a48b7a2025-08-20T04:03:07ZengNature PortfolioCommunications Biology2399-36422025-07-018111110.1038/s42003-025-08495-wConstruction and phenotypic classification of synthetic dual-pole Escherichia coli cellsTsung-Wei Chou0Ing-Jen Huang1Ya-Chu Hsu2Szu-Hua Wu3Jui-Chung Hong4Yu-Hao Ko5Ting-Ran Su6Jie-Hong Roland Jiang7Hsiao-Chun Huang8Institute of Molecular and Cellular Biology, National Taiwan UniversityInstitute of Molecular and Cellular Biology, National Taiwan UniversityInstitute of Molecular and Cellular Biology, National Taiwan UniversityInstitute of Molecular and Cellular Biology, National Taiwan UniversityInstitute of Molecular and Cellular Biology, National Taiwan UniversityGraduate Institute of Electronics Engineering, National Taiwan UniversityInstitute of Molecular and Cellular Biology, National Taiwan UniversityGraduate Institute of Electronics Engineering, National Taiwan UniversityInstitute of Molecular and Cellular Biology, National Taiwan UniversityAbstract Macromolecular complexes anchored at one cell pole have previously been utilized to achieve synthetic asymmetric cell division in Escherichia coli. In nature, however, robust asymmetric cell division often relies on gradients established between two opposing poles. Here, we present a second-generation synthetic genetic circuit that enables two distinct scaffolding proteins to localize at opposite poles in E. coli. By engineering a PodJ fragment as the second scaffold and integrating SpmX, a negative regulator of PodJ condensation, as a direct chimera with PopZ, we effectively confined PopZ and PodJ to opposite poles within E. coli. Additionally, we developed a neural network-based classification method to identify these synthetic dual-pole cells, further advancing the synthetic biology toolkit for programmable cell differentiation.https://doi.org/10.1038/s42003-025-08495-w
spellingShingle Tsung-Wei Chou
Ing-Jen Huang
Ya-Chu Hsu
Szu-Hua Wu
Jui-Chung Hong
Yu-Hao Ko
Ting-Ran Su
Jie-Hong Roland Jiang
Hsiao-Chun Huang
Construction and phenotypic classification of synthetic dual-pole Escherichia coli cells
Communications Biology
title Construction and phenotypic classification of synthetic dual-pole Escherichia coli cells
title_full Construction and phenotypic classification of synthetic dual-pole Escherichia coli cells
title_fullStr Construction and phenotypic classification of synthetic dual-pole Escherichia coli cells
title_full_unstemmed Construction and phenotypic classification of synthetic dual-pole Escherichia coli cells
title_short Construction and phenotypic classification of synthetic dual-pole Escherichia coli cells
title_sort construction and phenotypic classification of synthetic dual pole escherichia coli cells
url https://doi.org/10.1038/s42003-025-08495-w
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