A Framework for the Modular and Combinatorial Assembly of Synthetic Gene Circuits.

Details

Serval ID
serval:BIB_842B803713F3
Type
Article: article from journal or magazin.
Collection
Publications
Title
A Framework for the Modular and Combinatorial Assembly of Synthetic Gene Circuits.
Journal
ACS synthetic biology
Author(s)
Santos-Moreno J., Schaerli Y.
ISSN
2161-5063 (Electronic)
ISSN-L
2161-5063
Publication state
Published
Issued date
19/07/2019
Peer-reviewed
Oui
Volume
8
Number
7
Pages
1691-1697
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Synthetic gene circuits emerge from iterative design-build-test cycles. Most commonly, the time-limiting step is the circuit construction process. Here, we present a hierarchical cloning scheme based on the widespread Gibson assembly method and make the set of constructed plasmids freely available. Our two-step modular cloning scheme allows for simple, fast, efficient, and accurate assembly of gene circuits and combinatorial circuit libraries in Escherichia coli. The first step involves Gibson assembly of transcriptional units from constituent parts into individual intermediate plasmids. In the second step, these plasmids are digested with specific sets of restriction enzymes. The resulting flanking regions have overlaps that drive a second Gibson assembly into a single plasmid to yield the final circuit. This approach substantially reduces time and sequencing costs associated with gene circuit construction and allows for modular and combinatorial assembly of circuits. We demonstrate the usefulness of our framework by assembling a CRISPR-based double-inverter circuit and a combinatorial library of 3-node networks.
Keywords
DNA assembly, Gibson assembly, cloning, gene circuit, synthetic biology
Pubmed
Web of science
Create date
19/08/2019 13:36
Last modification date
21/08/2019 6:35
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