Robust and tunable signal processing in mammalian cells via engineered covalent modification cycles
Name
s41467-022-29338-w.pdf
Description
Published version
Size
1.99 MB
Format
Adobe PDF
Checksum (MD5)
b42549865fcf3a43b0a6b8927894926d
Author(s)
Jones, Ross D
Qian, Yili
Ilia, Katherine
Wang, Benjamin
Laub, Michael T
Del Vecchio, Domitilla
Weiss, Ron
Date Issued
2022
Journal
Nature Communications
Publisher
Springer Science and Business Media LLC
Citation
Jones, Ross D, Qian, Yili, Ilia, Katherine, Wang, Benjamin, Laub, Michael T et al. 2022. "Robust and tunable signal processing in mammalian cells via engineered covalent modification cycles." Nature Communications, 13 (1).
Version
Final published version
Abstract
AbstractEngineered signaling networks can impart cells with new functionalities useful for directing differentiation and actuating cellular therapies. For such applications, the engineered networks must be tunable, precisely regulate target gene expression, and be robust to perturbations within the complex context of mammalian cells. Here, we use bacterial two-component signaling proteins to develop synthetic phosphoregulation devices that exhibit these properties in mammalian cells. First, we engineer a synthetic covalent modification cycle based on kinase and phosphatase proteins derived from the bifunctional histidine kinase EnvZ, enabling analog tuning of gene expression via its response regulator OmpR. By regulating phosphatase expression with endogenous miRNAs, we demonstrate cell-type specific signaling responses and a new strategy for accurate cell type classification. Finally, we implement a tunable negative feedback controller via a small molecule-stabilized phosphatase, reducing output expression variance and mitigating the context-dependent effects of off-target regulation and resource competition. Our work lays the foundation for establishing tunable, precise, and robust control over cell behavior with synthetic signaling networks.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Terms of Use
Creative Commons Attribution 4.0 International license
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DOI of Published Version
https://doi.org/10.1038/S41467-022-29338-W
