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Synthetic genetic circuits to uncover and enforce the OCT4 trajectories of successful reprogramming of human fibroblasts
Reprogramming human fibroblasts to induced pluripotent stem cells (iPSCs) is inefficient, with heterogeneity among transcription factor (TF) trajectories driving divergent cell states. Nevertheless, the impact of TF dynamics on reprogramming efficiency remains uncharted. Here, we identify the succes...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9900859/ https://www.ncbi.nlm.nih.gov/pubmed/36747813 http://dx.doi.org/10.1101/2023.01.25.525529 |
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author | Ilia, Katherine Shakiba, Nika Bingham, Trevor Jones, Ross D. Kaminski, Michael M. Aravera, Eliezer Bruno, Simone Palacios, Sebastian Weiss, Ron Collins, James J. Del Vecchio, Domitilla Schlaeger, Thorsten M. |
author_facet | Ilia, Katherine Shakiba, Nika Bingham, Trevor Jones, Ross D. Kaminski, Michael M. Aravera, Eliezer Bruno, Simone Palacios, Sebastian Weiss, Ron Collins, James J. Del Vecchio, Domitilla Schlaeger, Thorsten M. |
author_sort | Ilia, Katherine |
collection | PubMed |
description | Reprogramming human fibroblasts to induced pluripotent stem cells (iPSCs) is inefficient, with heterogeneity among transcription factor (TF) trajectories driving divergent cell states. Nevertheless, the impact of TF dynamics on reprogramming efficiency remains uncharted. Here, we identify the successful reprogramming trajectories of the core pluripotency TF, OCT4, and design a genetic controller that enforces such trajectories with high precision. By combining a genetic circuit that generates a wide range of OCT4 trajectories with live-cell imaging, we track OCT4 trajectories with clonal resolution and find that a distinct constant OCT4 trajectory is required for colony formation. We then develop a synthetic genetic circuit that yields a tight OCT4 distribution around the identified trajectory and outperforms in terms of reprogramming efficiency other circuits that less accurately regulate OCT4. Our synthetic biology approach is generalizable for identifying and enforcing TF dynamics for cell fate programming applications. |
format | Online Article Text |
id | pubmed-9900859 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-99008592023-02-07 Synthetic genetic circuits to uncover and enforce the OCT4 trajectories of successful reprogramming of human fibroblasts Ilia, Katherine Shakiba, Nika Bingham, Trevor Jones, Ross D. Kaminski, Michael M. Aravera, Eliezer Bruno, Simone Palacios, Sebastian Weiss, Ron Collins, James J. Del Vecchio, Domitilla Schlaeger, Thorsten M. bioRxiv Article Reprogramming human fibroblasts to induced pluripotent stem cells (iPSCs) is inefficient, with heterogeneity among transcription factor (TF) trajectories driving divergent cell states. Nevertheless, the impact of TF dynamics on reprogramming efficiency remains uncharted. Here, we identify the successful reprogramming trajectories of the core pluripotency TF, OCT4, and design a genetic controller that enforces such trajectories with high precision. By combining a genetic circuit that generates a wide range of OCT4 trajectories with live-cell imaging, we track OCT4 trajectories with clonal resolution and find that a distinct constant OCT4 trajectory is required for colony formation. We then develop a synthetic genetic circuit that yields a tight OCT4 distribution around the identified trajectory and outperforms in terms of reprogramming efficiency other circuits that less accurately regulate OCT4. Our synthetic biology approach is generalizable for identifying and enforcing TF dynamics for cell fate programming applications. Cold Spring Harbor Laboratory 2023-01-25 /pmc/articles/PMC9900859/ /pubmed/36747813 http://dx.doi.org/10.1101/2023.01.25.525529 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator. |
spellingShingle | Article Ilia, Katherine Shakiba, Nika Bingham, Trevor Jones, Ross D. Kaminski, Michael M. Aravera, Eliezer Bruno, Simone Palacios, Sebastian Weiss, Ron Collins, James J. Del Vecchio, Domitilla Schlaeger, Thorsten M. Synthetic genetic circuits to uncover and enforce the OCT4 trajectories of successful reprogramming of human fibroblasts |
title | Synthetic genetic circuits to uncover and enforce the OCT4 trajectories of successful reprogramming of human fibroblasts |
title_full | Synthetic genetic circuits to uncover and enforce the OCT4 trajectories of successful reprogramming of human fibroblasts |
title_fullStr | Synthetic genetic circuits to uncover and enforce the OCT4 trajectories of successful reprogramming of human fibroblasts |
title_full_unstemmed | Synthetic genetic circuits to uncover and enforce the OCT4 trajectories of successful reprogramming of human fibroblasts |
title_short | Synthetic genetic circuits to uncover and enforce the OCT4 trajectories of successful reprogramming of human fibroblasts |
title_sort | synthetic genetic circuits to uncover and enforce the oct4 trajectories of successful reprogramming of human fibroblasts |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9900859/ https://www.ncbi.nlm.nih.gov/pubmed/36747813 http://dx.doi.org/10.1101/2023.01.25.525529 |
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