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...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2017-09-01
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| Series: | Cell Reports |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S221112471731152X |
| _version_ | 1818142460386213888 |
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| author | Toni A. Baeumler Ahmed Ashour Ahmed Tudor A. Fulga |
| author_facet | Toni A. Baeumler Ahmed Ashour Ahmed Tudor A. Fulga |
| author_sort | Toni A. Baeumler |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-11T11:16:08Z |
| format | Article |
| id | doaj.art-52f0672a227e47cc8af10d1cfe29b3e0 |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-12-11T11:16:08Z |
| publishDate | 2017-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-52f0672a227e47cc8af10d1cfe29b3e02022-12-22T01:09:19ZengElsevierCell Reports2211-12472017-09-0120112639265310.1016/j.celrep.2017.08.044Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric ReceptorsToni A. Baeumler0Ahmed Ashour Ahmed1Tudor A. Fulga2Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford OX3 9DS, UKOvarian Cancer Cell Laboratory, Weatherall Institute of Molecular Medicine, Nuffield Department of Obstetrics and Gynaecology, University of Oxford, Oxford OX3 9DU, UKWeatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford OX3 9DS, UKSynthetic 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.http://www.sciencedirect.com/science/article/pii/S221112471731152Xsynthetic receptorschimeric receptorsCRISPRdCas9-VP64split Cas9GPCRRTKgenome engineeringdCas9-synRtranscriptional programs |
| spellingShingle | Toni A. Baeumler Ahmed Ashour Ahmed Tudor A. Fulga Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors Cell Reports synthetic receptors chimeric receptors CRISPR dCas9-VP64 split Cas9 GPCR RTK genome engineering dCas9-synR transcriptional programs |
| 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 | synthetic receptors chimeric receptors CRISPR dCas9-VP64 split Cas9 GPCR RTK genome engineering dCas9-synR transcriptional programs |
| url | http://www.sciencedirect.com/science/article/pii/S221112471731152X |
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