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Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors

Synthetic receptors provide a powerful experimental tool for generation of designer cells capable of monitoring the environment, sensing specific input signals, and executing diverse custom response programs. To advance the promise of cellular engineering, we have developed a class of chimeric recep...

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Detalles Bibliográficos
Autores principales: Baeumler, Toni A., Ahmed, Ahmed Ashour, Fulga, Tudor A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5608971/
https://www.ncbi.nlm.nih.gov/pubmed/28903044
http://dx.doi.org/10.1016/j.celrep.2017.08.044
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author Baeumler, Toni A.
Ahmed, Ahmed Ashour
Fulga, Tudor A.
author_facet Baeumler, Toni A.
Ahmed, Ahmed Ashour
Fulga, Tudor A.
author_sort Baeumler, Toni A.
collection PubMed
description Synthetic receptors provide a powerful experimental tool for generation of designer cells capable of monitoring the environment, sensing specific input signals, and executing diverse custom response programs. To advance the promise of cellular engineering, we have developed a class of chimeric receptors that integrate a highly programmable and portable nuclease-deficient CRISPR/Cas9 (dCas9) signal transduction module. We demonstrate that the core dCas9 synthetic receptor (dCas9-synR) architecture can be readily adapted to various classes of native ectodomain scaffolds, linking their natural inputs with orthogonal output functions. Importantly, these receptors achieved stringent OFF/ON state transition characteristics, showed agonist-mediated dose-dependent activation, and could be programmed to couple specific disease markers with diverse, therapeutically relevant multi-gene expression circuits. The modular dCas9-synR platform developed here provides a generalizable blueprint for designing next generations of synthetic receptors, which will enable the implementation of highly complex combinatorial functions in cellular engineering.
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spelling pubmed-56089712017-09-29 Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors Baeumler, Toni A. Ahmed, Ahmed Ashour Fulga, Tudor A. Cell Rep Article Synthetic receptors provide a powerful experimental tool for generation of designer cells capable of monitoring the environment, sensing specific input signals, and executing diverse custom response programs. To advance the promise of cellular engineering, we have developed a class of chimeric receptors that integrate a highly programmable and portable nuclease-deficient CRISPR/Cas9 (dCas9) signal transduction module. We demonstrate that the core dCas9 synthetic receptor (dCas9-synR) architecture can be readily adapted to various classes of native ectodomain scaffolds, linking their natural inputs with orthogonal output functions. Importantly, these receptors achieved stringent OFF/ON state transition characteristics, showed agonist-mediated dose-dependent activation, and could be programmed to couple specific disease markers with diverse, therapeutically relevant multi-gene expression circuits. The modular dCas9-synR platform developed here provides a generalizable blueprint for designing next generations of synthetic receptors, which will enable the implementation of highly complex combinatorial functions in cellular engineering. Cell Press 2017-09-12 /pmc/articles/PMC5608971/ /pubmed/28903044 http://dx.doi.org/10.1016/j.celrep.2017.08.044 Text en Crown Copyright © 2017 Published by Elsevier Inc. All rights reserved. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Baeumler, Toni A.
Ahmed, Ahmed Ashour
Fulga, Tudor A.
Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors
title Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors
title_full Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors
title_fullStr Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors
title_full_unstemmed Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors
title_short Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors
title_sort engineering synthetic signaling pathways with programmable dcas9-based chimeric receptors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5608971/
https://www.ncbi.nlm.nih.gov/pubmed/28903044
http://dx.doi.org/10.1016/j.celrep.2017.08.044
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