Combinatorial metabolic pathway assembly approaches and toolkits for modular assembly
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Published version
Author(s)
Young, Rosanna
Haines, Matthew
Storch, Marko
Freemont, Paul S
Type
Journal Article
Abstract
Synthetic Biology is a rapidly growing interdisciplinary field that is primarily built upon foundational advances in molecular biology combined with engineering design principles such as modularity and interoperability. The field considers living systems as programmable at the genetic level and has been defined by the development of new platform technologies and methodological advances. A key concept driving the field is the Design-Build-Test-Learn cycle which provides a systematic framework for building new biological systems. One major application area for synthetic biology is biosynthetic pathway engineering that requires the modular assembly of different genetic regulatory elements and biosynthetic enzymes. In this review we provide an overview of modular DNA assembly and describe and compare the plethora of in vitro and in vivo assembly methods for combinatorial pathway engineering. Considerations for part design and methods for enzyme balancing are also presented, and we briefly discuss alternatives to intracellular pathway assembly including microbial consortia and cell-free systems for biosynthesis. Finally, we describe computational tools and automation for pathway design and assembly and argue that a deeper understanding of the many different variables of genetic design, pathway regulation and cellular metabolism will allow more predictive pathway design and engineering.
Date Issued
2021-01-01
Date Acceptance
2020-12-03
Citation
Metabolic Engineering, 2021, 63, pp.81-101
ISSN
1096-7176
Publisher
Elsevier
Start Page
81
End Page
101
Journal / Book Title
Metabolic Engineering
Volume
63
Copyright Statement
Crown Copyright © 2020 Published by Elsevier Inc. on behalf of International Metabolic Engineering Society. This is an open access article under theCC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/33301873
PII: S1096-7176(20)30185-3
Subjects
Chassis
Cloning
DNA assembly
Pathway tuning
Synthetic biology
Publication Status
Published
Coverage Spatial
Belgium
Date Publish Online
2020-12-07