Multiplex Expression Cassette Assembly: A flexible and versatile method for building complex genetic circuits in conventional vectors Article Swipe
Xun Jiang
,
Zhuoxiang Zhang
,
Xiuming Wu
,
Changmei Li
,
Xuan Sun
,
Yiting Li
,
Aixia Chang
,
Aiguo Yang
,
Changqing Yang
·
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.1111/pbi.14454
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.1111/pbi.14454
Summary The manipulation of multiple transcription units for simultaneous and coordinated expression is not only key to building complex genetic circuits to accomplish diverse functions in synthetic biology, but is also important in crop breeding for significantly improved productivity and overall performance. However, building constructs with multiple independent transcription units for fine‐tuned and coordinated regulation is complicated and time‐consuming. Here, we introduce the Multiplex Expression Cassette Assembly (MECA) method, which modifies canonical vectors compatible with Golden Gate Assembly, and then uses them to produce multi‐cassette constructs. By embedding the junction syntax in primers that are used to amplify functional elements, MECA is able to make complex constructs using only one intermediate vector and one destination vector via two rounds of one‐pot Golden Gate assembly reactions, without the need for dedicated vectors and a coherent library of standardized modules. As a proof‐of‐concept, we modified eukaryotic and prokaryotic expression vectors to generate constructs for transient expression of green fluorescent protein and β‐glucuronidase in Nicotiana benthamiana , genome editing to block monoterpene metabolism in tomato glandular trichomes, production of betanin in tobacco and synthesis of β‐carotene in Escherichia coli . Additionally, we engineered the stable production of thymol and carvacrol, bioactive compounds from Lamiaceae family plants, in glandular trichomes of tobacco. These results demonstrate that MECA is a flexible, efficient and versatile method for building complex genetic circuits, which will not only play a critical role in plant synthetic biology, but also facilitate improving agronomic traits and pyramiding traits for the development of next‐generation elite crops.
Related Topics
Concepts
Biology
Synthetic biology
Phytoene desaturase
Expression cassette
Computational biology
Nicotiana benthamiana
Genetics
Vector (molecular biology)
Gene
Recombinant DNA
Biosynthesis
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1111/pbi.14454
- OA Status
- gold
- Cited By
- 4
- References
- 95
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4401821561
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4401821561Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1111/pbi.14454Digital Object Identifier
- Title
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Multiplex Expression Cassette Assembly: A flexible and versatile method for building complex genetic circuits in conventional vectorsWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2024Year of publication
- Publication date
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2024-08-23Full publication date if available
- Authors
-
Xun Jiang, Zhuoxiang Zhang, Xiuming Wu, Changmei Li, Xuan Sun, Yiting Li, Aixia Chang, Aiguo Yang, Changqing YangList of authors in order
- Landing page
-
https://doi.org/10.1111/pbi.14454Publisher landing page
- Open access
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YesWhether a free full text is available
- OA status
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goldOpen access status per OpenAlex
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https://doi.org/10.1111/pbi.14454Direct OA link when available
- Concepts
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Biology, Synthetic biology, Phytoene desaturase, Expression cassette, Computational biology, Nicotiana benthamiana, Genetics, Vector (molecular biology), Gene, Recombinant DNA, BiosynthesisTop concepts (fields/topics) attached by OpenAlex
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4Total citation count in OpenAlex
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2025: 4Per-year citation counts (last 5 years)
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10Other works algorithmically related by OpenAlex
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