Compact engineered human mechanosensitive transactivation modules enable potent and versatile synthetic transcriptional control

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dc.citation.firstpage1716
dc.citation.issueNumber11
dc.citation.journalTitleNature Methods
dc.citation.lastpage1728
dc.citation.volumeNumber20
dc.contributor.authorMahata, Barun
dc.contributor.authorCabrera, Alan
dc.contributor.authorBrenner, Daniel A.
dc.contributor.authorGuerra-Resendez, Rosa Selenia
dc.contributor.authorLi, Jing
dc.contributor.authorGoell, Jacob
dc.contributor.authorWang, Kaiyuan
dc.contributor.authorGuo, Yannie
dc.contributor.authorEscobar, Mario
dc.contributor.authorParthasarathy, Abinand Krishna
dc.contributor.authorSzadowski, Hailey
dc.contributor.authorBedford, Guy
dc.contributor.authorReed, Daniel R.
dc.contributor.authorKim, Sunghwan
dc.contributor.authorHilton, Isaac B.
dc.date.accessioned2024-05-03T15:51:20Z
dc.date.available2024-05-03T15:51:20Z
dc.date.issued2023
dc.description.abstractEngineered transactivation domains (TADs) combined with programmable DNA binding platforms have revolutionized synthetic transcriptional control. Despite recent progress in programmable CRISPR–Cas-based transactivation (CRISPRa) technologies, the TADs used in these systems often contain poorly tolerated elements and/or are prohibitively large for many applications. Here, we defined and optimized minimal TADs built from human mechanosensitive transcription factors. We used these components to construct potent and compact multipartite transactivation modules (MSN, NMS and eN3x9) and to build the CRISPR–dCas9 recruited enhanced activation module (CRISPR-DREAM) platform. We found that CRISPR-DREAM was specific and robust across mammalian cell types, and efficiently stimulated transcription from diverse regulatory loci. We also showed that MSN and NMS were portable across Type I, II and V CRISPR systems, transcription activator-like effectors and zinc finger proteins. Further, as proofs of concept, we used dCas9-NMS to efficiently reprogram human fibroblasts into induced pluripotent stem cells and demonstrated that mechanosensitive transcription factor TADs are efficacious and well tolerated in therapeutically important primary human cell types. Finally, we leveraged the compact and potent features of these engineered TADs to build dual and all-in-one CRISPRa AAV systems. Altogether, these compact human TADs, fusion modules and delivery architectures should be valuable for synthetic transcriptional control in biomedical applications.
dc.identifier.citationMahata, B., Cabrera, A., Brenner, D. A., Guerra-Resendez, R. S., Li, J., Goell, J., Wang, K., Guo, Y., Escobar, M., Parthasarathy, A. K., Szadowski, H., Bedford, G., Reed, D. R., Kim, S., & Hilton, I. B. (2023). Compact engineered human mechanosensitive transactivation modules enable potent and versatile synthetic transcriptional control. Nature Methods, 20(11), 1716–1728. https://doi.org/10.1038/s41592-023-02036-1
dc.identifier.digitals41592-023-02036-1
dc.identifier.doihttps://doi.org/10.1038/s41592-023-02036-1
dc.identifier.urihttps://hdl.handle.net/1911/115622
dc.language.isoeng
dc.publisherSpringer Nature
dc.rightsExcept where otherwise noted, this work is licensed under a Creative Commons Attribution (CC BY) license. Permission to reuse, publish, or reproduce the work beyond the terms of the license or beyond the bounds of fair use or other exemptions to copyright law must be obtained from the copyright holder.
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.titleCompact engineered human mechanosensitive transactivation modules enable potent and versatile synthetic transcriptional control
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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