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  A Step-by-Step Protocol for COMPASS, a Synthetic Biology Tool for Combinatorial Gene Assembly

Naseri, G., & Mueller-Roeber, B. (2020). A Step-by-Step Protocol for COMPASS, a Synthetic Biology Tool for Combinatorial Gene Assembly. In S. Chandran, & K. W. George (Eds.), DNA Cloning and Assembly: Methods and Protocols (pp. 277-303). New York, NY: Springer US.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0006-E58E-8 Version Permalink: http://hdl.handle.net/21.11116/0000-0006-E58F-7
Genre: Book Chapter

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 Creators:
Naseri, Gita1, Author
Mueller-Roeber, B.2, Author              
Affiliations:
1External Organizations, ou_persistent22              
2Plant Signalling, Cooperative Research Groups, Max Planck Institute of Molecular Plant Physiology, Max Planck Society, ou_1753311              

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 Abstract: For industry-scale production of high-value chemicals in microbial cell factories, the elimination of metabolic flux imbalances is a critical aspect. However, a priori knowledge about the genetic design of optimal production pathways is typically not available. COMPASS, COMbinatorial Pathway ASSembly, is a rapid cloning method for the balanced expression of multiple genes in biochemical pathways. The method generates thousands of individual DNA constructs in modular, parallel, and high-throughput manner. COMPASS employs inducible artificial transcription factors derived from plant (Arabidopsis thaliana) regulators to control the expression of pathway genes in yeast (Saccharomyces cerevisiae). It utilizes homologous recombination for parts assembly and employs a positive selection scheme to identify correctly assembled pathway variants after both in vivo and in vitro recombination. Finally, COMPASS is equipped with a CRISPR/Cas9 genome modification system allowing for the one-step multilocus integration of genes. Although COMPASS was initially developed for pathway engineering, it can equally be employed for balancing gene expression in other synthetic biology projects.

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Language(s): eng - English
 Dates: 2020
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: -
 Identifiers: DOI: 10.1007/978-1-0716-0908-8_16
BibTex Citekey: Naseri2020
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Title: DNA Cloning and Assembly: Methods and Protocols
Source Genre: Book
 Creator(s):
Chandran, Sunil, Editor
George, Kevin W., Editor
Affiliations:
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Publ. Info: New York, NY : Springer US
Pages: - Volume / Issue: - Sequence Number: - Start / End Page: 277 - 303 Identifier: ISBN: 978-1-0716-0908-8