Modular cloning by Golden Gate assembly and possible application in pathway design

Raguz Nakic, Zrinka; Peters, Christin

doi:10.2533/chimia.2023.437

Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-29951

Publication type:	Article in scientific journal
Type of review:	Peer review (publication)
Title:	Modular cloning by Golden Gate assembly and possible application in pathway design
Authors:	Raguz Nakic, Zrinka Peters, Christin
et. al:	No
DOI:	10.2533/chimia.2023.437 10.21256/zhaw-29951
Published in:	Chimia
Volume(Issue):	77
Issue:	6
Page(s):	437
Pages to:	441
Issue Date:	28-Jun-2023
Publisher / Ed. Institution:	Swiss Chemical Society
ISSN:	0009-4293
Language:	English
Subjects:	Expression control; Golden Gate assembly; Modular cloning; Pathway design; Binding sites; Promoter regions, genetic; Cloning, molecular; High-throughput screening assay; Synthetic biology
Subject (DDC):	572: Biochemistry 660.6: Biotechnology
Abstract:	Preparation of expression vectors using conventional cloning strategies is laborious and not suitable for the design of metabolic pathways or enzyme cascades, which usually requires the preparation of a vector library to identify productive clones. Recently, Modular Cloning as a novel cloning technique in synthetic biology has been developed. Modular Cloning relies on Golden Gate assembly and supports preparation of individual expression vectors in one-step and one-pot reactions, thus allowing rapid generation of vector libraries. A number of Modular Cloning toolkits for specific applications has been established, providing a collection of distinct genetic elements such as promoters, ribosome binding sites and tags, that can be combined individually in one-step using defined fusion sites. Modular Cloning has been successfully applied to generate various strains for producing value-added compounds. This was achieved by orchestrating complex pathways involving up to 20 enzymes. Due to the novelty of the genetic approach, industrial applications are still rare. In addition, some applications are limited due to the lack of high-throughput screening methods. This shifts the bottleneck from library preparation to screening capacity and needs to be addressed by future developments to pave the path for the establishment of Modular Cloning in industrial applications.
URI:	https://digitalcollection.zhaw.ch/handle/11475/29951
Fulltext version:	Published version
License (according to publishing contract):	CC BY 4.0: Attribution 4.0 International
Departement:	Life Sciences and Facility Management
Organisational Unit:	Institute of Chemistry and Biotechnology (ICBT)
Published as part of the ZHAW project:	Plasmid-Toolbox für Enzymkaskaden
Appears in collections:	Publikationen Life Sciences und Facility Management

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Raguz Nakic, Z., & Peters, C. (2023). Modular cloning by Golden Gate assembly and possible application in pathway design. Chimia, 77(6), 437–441. https://doi.org/10.2533/chimia.2023.437

Raguz Nakic, Z. and Peters, C. (2023) ‘Modular cloning by Golden Gate assembly and possible application in pathway design’, Chimia, 77(6), pp. 437–441. Available at: https://doi.org/10.2533/chimia.2023.437.

Z. Raguz Nakic and C. Peters, “Modular cloning by Golden Gate assembly and possible application in pathway design,” Chimia, vol. 77, no. 6, pp. 437–441, Jun. 2023, doi: 10.2533/chimia.2023.437.

RAGUZ NAKIC, Zrinka und Christin PETERS, 2023. Modular cloning by Golden Gate assembly and possible application in pathway design. Chimia. 28 Juni 2023. Bd. 77, Nr. 6, S. 437–441. DOI 10.2533/chimia.2023.437

Raguz Nakic, Zrinka, and Christin Peters. 2023. “Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design.” Chimia 77 (6): 437–41. https://doi.org/10.2533/chimia.2023.437.

Raguz Nakic, Zrinka, and Christin Peters. “Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design.” Chimia, vol. 77, no. 6, June 2023, pp. 437–41, https://doi.org/10.2533/chimia.2023.437.