A modular synthetic biology toolkit unlocks metabolic engineering of the industrially relevant alga Nannochloropsis.

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Autores principales: Guo, Yutan, Li, Zhixiong, Wang, Hao, Zheng, Jie, Liang, Zhiwei, Sun, Han, Zhou, Wenguang, Liu, Jin
Formato: Artículo científico
Lenguaje:en
Publicado: Advanced biotechnology 2026
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author Guo, Yutan
Li, Zhixiong
Wang, Hao
Zheng, Jie
Liang, Zhiwei
Sun, Han
Zhou, Wenguang
Liu, Jin
author_facet Guo, Yutan
Li, Zhixiong
Wang, Hao
Zheng, Jie
Liang, Zhiwei
Sun, Han
Zhou, Wenguang
Liu, Jin
Guo, Yutan
Li, Zhixiong
Wang, Hao
Zheng, Jie
Liang, Zhiwei
Sun, Han
Zhou, Wenguang
Liu, Jin
collection PubMed - marine biology
contents A modular synthetic biology toolkit unlocks metabolic engineering of the industrially relevant alga Nannochloropsis. Guo, Yutan Li, Zhixiong Wang, Hao Zheng, Jie Liang, Zhiwei Sun, Han Zhou, Wenguang Liu, Jin Nannochloropsis is an industrially relevant marine microalga with exceptional potential as a chassis for sunlight-driven CO valorization. However, its broad application in synthetic biology has been constrained by the lack of a standardized and modular genetic toolbox. Here, we report the development of a comprehensive Modular Cloning (MoClo) toolkit for Nannochloropsis, based on Golden Gate assembly and a standard syntax. The toolkit comprises 91 domesticated genetic parts spanning promoters, signal peptides, selectable markers, reporter genes, tags and terminators. A large subset of these parts, including several not previously evaluated in Nannochloropsis, was functionally validated, enabling convenient and reliable transformant selection, immunodetection, and subcellular localization. To demonstrate the utility of the toolkit for multi-gene pathway engineering, modularly assembled keto-carotenoid biosynthetic pathways were introduced into Nannochloropsis, leading to substantial accumulation of canthaxanthin (4.5 mg g) or astaxanthin (2.8 mg g). Collectively, this flexible and expandable MoClo toolkit establishes a standardized foundation for synthetic biology in Nannochloropsis, enables rapid design-build-test cycles for multi-gene constructs, and advances the use of industrial microalga for sustainable, CO-based production of value-added biochemicals.
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publishDate 2026
publisher Advanced biotechnology
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spellingShingle A modular synthetic biology toolkit unlocks metabolic engineering of the industrially relevant alga Nannochloropsis.
Guo, Yutan
Li, Zhixiong
Wang, Hao
Zheng, Jie
Liang, Zhiwei
Sun, Han
Zhou, Wenguang
Liu, Jin
A modular synthetic biology toolkit unlocks metabolic engineering of the industrially relevant alga Nannochloropsis. Guo, Yutan Li, Zhixiong Wang, Hao Zheng, Jie Liang, Zhiwei Sun, Han Zhou, Wenguang Liu, Jin Nannochloropsis is an industrially relevant marine microalga with exceptional potential as a chassis for sunlight-driven CO valorization. However, its broad application in synthetic biology has been constrained by the lack of a standardized and modular genetic toolbox. Here, we report the development of a comprehensive Modular Cloning (MoClo) toolkit for Nannochloropsis, based on Golden Gate assembly and a standard syntax. The toolkit comprises 91 domesticated genetic parts spanning promoters, signal peptides, selectable markers, reporter genes, tags and terminators. A large subset of these parts, including several not previously evaluated in Nannochloropsis, was functionally validated, enabling convenient and reliable transformant selection, immunodetection, and subcellular localization. To demonstrate the utility of the toolkit for multi-gene pathway engineering, modularly assembled keto-carotenoid biosynthetic pathways were introduced into Nannochloropsis, leading to substantial accumulation of canthaxanthin (4.5 mg g) or astaxanthin (2.8 mg g). Collectively, this flexible and expandable MoClo toolkit establishes a standardized foundation for synthetic biology in Nannochloropsis, enables rapid design-build-test cycles for multi-gene constructs, and advances the use of industrial microalga for sustainable, CO-based production of value-added biochemicals.
title A modular synthetic biology toolkit unlocks metabolic engineering of the industrially relevant alga Nannochloropsis.
url https://pubmed.ncbi.nlm.nih.gov/41632226/