Fluorescein-based SynNotch adaptors for regulating gene expression responses to diverse extracellular and matrix-based cues
Abstract Synthetic Notch (SynNotch) receptors function like natural Notch proteins and can be used to install customized sense-and-respond capabilities into mammalian cells. Here, we introduce an adaptor-based strategy for regulating SynNotch activity via fluorescein isomers and analogs. Using an op...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56148-7 |
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| _version_ | 1832585606934298624 |
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| author | Jeremy C. Tran Christopher J. Kuffner Alexander M. Marzilli Ryan Emily Miller Zachary E. Silfen Jeffrey B. McMahan D. Christopher Sloas Christopher S. Chen John T. Ngo |
| author_facet | Jeremy C. Tran Christopher J. Kuffner Alexander M. Marzilli Ryan Emily Miller Zachary E. Silfen Jeffrey B. McMahan D. Christopher Sloas Christopher S. Chen John T. Ngo |
| author_sort | Jeremy C. Tran |
| collection | DOAJ |
| description | Abstract Synthetic Notch (SynNotch) receptors function like natural Notch proteins and can be used to install customized sense-and-respond capabilities into mammalian cells. Here, we introduce an adaptor-based strategy for regulating SynNotch activity via fluorescein isomers and analogs. Using an optimized fluorescein-binding SynNotch receptor, we describe ways to chemically control SynNotch signaling, including an approach based on a bio-orthogonal chemical ligation and a spatially controllable strategy via the photo-patterned uncaging of an o-nitrobenzyl-caged fluorescein conjugate. We further show that fluorescein-conjugated extracellular matrix (ECM)-binding peptides can be used to regulate SynNotch activity depending on the folding state of collagen-based ECM networks. To demonstrate the utility of these tools, we apply them to activate dose-dependent gene expression responses and to induce myogenic-like phenotypes in multipotent fibroblasts with spatiotemporal and microenvironmental control. Overall, we introduce an optimized fluorescein-binding SynNotch as a versatile tool for regulating transcriptional responses to ligands based on the clinically-approved fluorescein dye. |
| format | Article |
| id | doaj-art-f2e26143b187447e822842ebbcc0e10b |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-f2e26143b187447e822842ebbcc0e10b2025-01-26T12:41:18ZengNature PortfolioNature Communications2041-17232025-01-0116111710.1038/s41467-025-56148-7Fluorescein-based SynNotch adaptors for regulating gene expression responses to diverse extracellular and matrix-based cuesJeremy C. Tran0Christopher J. Kuffner1Alexander M. Marzilli2Ryan Emily Miller3Zachary E. Silfen4Jeffrey B. McMahan5D. Christopher Sloas6Christopher S. Chen7John T. Ngo8Department of Biomedical Engineering, Boston UniversityDepartment of Biomedical Engineering, Boston UniversityDepartment of Biomedical Engineering, Boston UniversityDepartment of Biomedical Engineering, Boston UniversityDepartment of Biomedical Engineering, Boston UniversityDepartment of Biomedical Engineering, Boston UniversityDepartment of Biomedical Engineering, Boston UniversityDepartment of Biomedical Engineering, Boston UniversityDepartment of Biomedical Engineering, Boston UniversityAbstract Synthetic Notch (SynNotch) receptors function like natural Notch proteins and can be used to install customized sense-and-respond capabilities into mammalian cells. Here, we introduce an adaptor-based strategy for regulating SynNotch activity via fluorescein isomers and analogs. Using an optimized fluorescein-binding SynNotch receptor, we describe ways to chemically control SynNotch signaling, including an approach based on a bio-orthogonal chemical ligation and a spatially controllable strategy via the photo-patterned uncaging of an o-nitrobenzyl-caged fluorescein conjugate. We further show that fluorescein-conjugated extracellular matrix (ECM)-binding peptides can be used to regulate SynNotch activity depending on the folding state of collagen-based ECM networks. To demonstrate the utility of these tools, we apply them to activate dose-dependent gene expression responses and to induce myogenic-like phenotypes in multipotent fibroblasts with spatiotemporal and microenvironmental control. Overall, we introduce an optimized fluorescein-binding SynNotch as a versatile tool for regulating transcriptional responses to ligands based on the clinically-approved fluorescein dye.https://doi.org/10.1038/s41467-025-56148-7 |
| spellingShingle | Jeremy C. Tran Christopher J. Kuffner Alexander M. Marzilli Ryan Emily Miller Zachary E. Silfen Jeffrey B. McMahan D. Christopher Sloas Christopher S. Chen John T. Ngo Fluorescein-based SynNotch adaptors for regulating gene expression responses to diverse extracellular and matrix-based cues Nature Communications |
| title | Fluorescein-based SynNotch adaptors for regulating gene expression responses to diverse extracellular and matrix-based cues |
| title_full | Fluorescein-based SynNotch adaptors for regulating gene expression responses to diverse extracellular and matrix-based cues |
| title_fullStr | Fluorescein-based SynNotch adaptors for regulating gene expression responses to diverse extracellular and matrix-based cues |
| title_full_unstemmed | Fluorescein-based SynNotch adaptors for regulating gene expression responses to diverse extracellular and matrix-based cues |
| title_short | Fluorescein-based SynNotch adaptors for regulating gene expression responses to diverse extracellular and matrix-based cues |
| title_sort | fluorescein based synnotch adaptors for regulating gene expression responses to diverse extracellular and matrix based cues |
| url | https://doi.org/10.1038/s41467-025-56148-7 |
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