Self-organization of embryonic stem cells into a reproducible embryo model through epigenome editing.
| العنوان: | Self-organization of embryonic stem cells into a reproducible embryo model through epigenome editing. |
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| المؤلفون: | Lodewijk GA; Department of Biomolecular Engineering, University of California, Santa Cruz, CA.; Genomics Institute, University of California, Santa Cruz, CA.; Institute for The Biology of Stem Cells, University of California, Santa Cruz, CA.; Equal contribution to this work., Kozuki S; Department of Biomolecular Engineering, University of California, Santa Cruz, CA.; Genomics Institute, University of California, Santa Cruz, CA.; Institute for The Biology of Stem Cells, University of California, Santa Cruz, CA.; Equal contribution to this work., Han C; Department of Biomolecular Engineering, University of California, Santa Cruz, CA.; Genomics Institute, University of California, Santa Cruz, CA.; Institute for The Biology of Stem Cells, University of California, Santa Cruz, CA., Topacio BR; Department of Biomolecular Engineering, University of California, Santa Cruz, CA.; Genomics Institute, University of California, Santa Cruz, CA.; Institute for The Biology of Stem Cells, University of California, Santa Cruz, CA., Zargari A; Department of Electrical and Computer Engineering, University of California, Santa Cruz, CA., Lee S; Department of Biomolecular Engineering, University of California, Santa Cruz, CA.; Genomics Institute, University of California, Santa Cruz, CA.; Institute for The Biology of Stem Cells, University of California, Santa Cruz, CA., Knight G; Neurosetta LLC, Madison, WI.; Wisconsin Institute for Discovery, Madison, WI., Ashton R; Neurosetta LLC, Madison, WI.; Wisconsin Institute for Discovery, Madison, WI.; Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI., Qi LS; Department of Bioengineering, Stanford University, Stanford, CA.; Sarafan ChEM-H, Stanford University, Stanford, CA.; Chan Zuckerberg Biohub - San Francisco, San Francisco, CA., Shariati SA; Department of Biomolecular Engineering, University of California, Santa Cruz, CA.; Genomics Institute, University of California, Santa Cruz, CA.; Institute for The Biology of Stem Cells, University of California, Santa Cruz, CA. |
| المصدر: | BioRxiv : the preprint server for biology [bioRxiv] 2024 Mar 10. Date of Electronic Publication: 2024 Mar 10. |
| نوع المنشور: | Preprint |
| اللغة: | English |
| بيانات الدورية: | Country of Publication: United States NLM ID: 101680187 Publication Model: Electronic Cited Medium: Internet NLM ISO Abbreviation: bioRxiv Subsets: PubMed not MEDLINE |
| مستخلص: | Embryonic stem cells (ESCs) can self-organize in vitro into developmental patterns with spatial organization and molecular similarity to that of early embryonic stages. This self-organization of ESCs requires transmission of signaling cues, via addition of small molecule chemicals or recombinant proteins, to induce distinct embryonic cellular fates and subsequent assembly into structures that can mimic aspects of early embryonic development. During natural embryonic development, different embryonic cell types co-develop together, where each cell type expresses specific fate-inducing transcription factors through activation of non-coding regulatory elements and interactions with neighboring cells. However, previous studies have not fully explored the possibility of engineering endogenous regulatory elements to shape self-organization of ESCs into spatially-ordered embryo models. Here, we hypothesized that cell-intrinsic activation of a minimum number of such endogenous regulatory elements is sufficient to self-organize ESCs into early embryonic models. Our results show that CRISPR-based activation (CRISPRa) of only two endogenous regulatory elements in the genome of pluripotent stem cells is sufficient to generate embryonic patterns that show spatial and molecular resemblance to that of pre-gastrulation mouse embryonic development. Quantitative single-cell live fluorescent imaging showed that the emergence of spatially-ordered embryonic patterns happens through the intrinsic induction of cell fate that leads to an orchestrated collective cellular motion. Based on these results, we propose a straightforward approach to efficiently form 3D embryo models through intrinsic CRISPRa-based epigenome editing and independent of external signaling cues. CRISPRa-Programmed Embryo Models (CPEMs) show highly consistent composition of major embryonic cell types that are spatially-organized, with nearly 80% of the structures forming an embryonic cavity. Single cell transcriptomics confirmed the presence of main embryonic cell types in CPEMs with transcriptional similarity to pre-gastrulation mouse embryos and revealed novel signaling communication links between different embryonic cell types. Our findings offer a programmable embryo model and demonstrate that minimum intrinsic epigenome editing is sufficient to self-organize ESCs into highly consistent pre-gastrulation embryo models. Competing Interests: Conflict of Interest G.K and R.A are co-founder of Neurosetta, which is focused on commercializing the RosetteArray platform. |
| معلومات مُعتمدة: | K12 GM139185 United States GM NIGMS NIH HHS; R00 GM126027 United States GM NIGMS NIH HHS; K99 GM126027 United States GM NIGMS NIH HHS; R42 ES033912 United States ES NIEHS NIH HHS; R35 GM147395 United States GM NIGMS NIH HHS |
| تواريخ الأحداث: | Date Created: 20240318 Latest Revision: 20240325 |
| رمز التحديث: | 20240325 |
| مُعرف محوري في PubMed: | PMC10942404 |
| DOI: | 10.1101/2024.03.05.583597 |
| PMID: | 38496557 |
| قاعدة البيانات: | MEDLINE |
| DOI: | 10.1101/2024.03.05.583597 |
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