Fiche du document
19 juillet 2019
- doi: 10.1021/acssynbio.9b00174
- issn: 2161-5063
Ce document est lié à :
info:eu-repo/semantics/altIdentifier/doi/10.1021/acssynbio.9b00174
Ce document est lié à :
info:eu-repo/semantics/altIdentifier/pmid/31185158
Ce document est lié à :
info:eu-repo/semantics/altIdentifier/eissn/2161-5063
Ce document est lié à :
info:eu-repo/semantics/altIdentifier/urn/urn:nbn:ch:serval-BIB_842B803713F37
info:eu-repo/semantics/openAccess , Copying allowed only for non-profit organizations , https://serval.unil.ch/disclaimer
Sujets proches
Paragenes Circuits, Gene Gene modules Gene networks Genetic regulatory networks Transcriptional networks Networks, Transcriptional Regulatory networks, Gene GRNs (Gene regulatory networks) Gene circuits Modules, Gene Networks, Gene regulatoryCiter ce document
J. Santos-Moreno et al., « A Framework for the Modular and Combinatorial Assembly of Synthetic Gene Circuits. », Serveur académique Lausannois, ID : 10.1021/acssynbio.9b00174
Métriques
Partage / Export
Résumé
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.