Modular Extracellular sensor architecture for engineering mammalian cell-based devices

Nichole M. Daringer, Rachel M. Dudek, Kelly A. Schwarz, Joshua N. Leonard*

*Corresponding author for this work

Research output: Contribution to journalArticle

49 Scopus citations

Abstract

Engineering mammalian cell-based devices that monitor and therapeutically modulate human physiology is a promising and emerging frontier in clinical synthetic biology. However, realizing this vision will require new technologies enabling engineered circuitry to sense and respond to physiologically relevant cues. No existing technology enables an engineered cell to sense exclusively extracellular ligands, including proteins and pathogens, without relying upon native cellular receptors or signal transduction pathways that may be subject to crosstalk with native cellular components. To address this need, we here report a technology we term a Modular Extracellular Sensor Architecture (MESA). This self-contained receptor and signal transduction platform is maximally orthogonal to native cellular processes and comprises independent, tunable protein modules that enable performance optimization and straightforward engineering of novel MESA that recognize novel ligands. We demonstrate ligand-inducible activation of MESA signaling, optimization of receptor performance using design-based approaches, and generation of MESA biosensors that produce outputs in the form of either transcriptional regulation or transcription-independent reconstitution of enzymatic activity. This systematic, quantitative platform characterization provides a framework for engineering MESA to recognize novel ligands and for integrating these sensors into diverse mammalian synthetic biology applications.

Original languageEnglish (US)
Pages (from-to)892-902
Number of pages11
JournalACS synthetic biology
Volume3
Issue number12
DOIs
StatePublished - Dec 19 2014

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Keywords

  • biosensor
  • cell therapy
  • mammalian synthetic biology
  • receptor engineering

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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