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Engineering of an Optogenetic T Cell Receptor Compatible with Fluorescence-Based Readouts.

Vincent IdsteinAnna K EhretO Sascha YousefiWolfgang W A Schamel
Published in: ACS synthetic biology (2023)
Optogenetics offers a set of tools for the precise manipulation of signaling pathways. Here we exploit optogenetics to experimentally change the kinetics of protein-protein interactions on demand. We had developed a system in which the interaction of a modified T cell receptor (TCR) with an engineered ligand can be controlled by light. The ligand was the plant photoreceptor phytochrome B (PhyB) and the TCR included a TCRβ chain fused to GFP and a mutated PhyB-interacting factor (PIF S ), resulting in the GFP-PIF S -TCR. We failed to engineer a nonfluorescent PIF S -fused TCR, since PIF S did not bind to PhyB when omitting GFP. Here we tested nine different versions of PIF S -fused TCRs. We found that the SNAP-PIF S -TCR was expressed well on the surface, bound to PhyB, and subsequently elicited activation signals. This receptor could be combined with a GFP reporter system in which the expression of GFP is driven by the transcription factor NF-AT.
Keyphrases
  • regulatory t cells
  • signaling pathway
  • transcription factor
  • dendritic cells
  • binding protein
  • poor prognosis
  • lps induced
  • immune response
  • long non coding rna
  • toll like receptor