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Programming cellular condensates for living materials using an intrinsically disordered protein display platform.

Rong Chang1, Hann X Tu1, Hongyu Ma2

  • 1Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115.

Proceedings of the National Academy of Sciences of the United States of America
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We developed a new platform for displaying intrinsically disordered proteins (IDPs) on bacteria, enabling programmable self-organization and the creation of dynamic living materials with tunable properties.

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cell surface displayintrinsically disordered proteinliving materialsmulticellular patterning

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Area of Science:

  • Synthetic Biology
  • Biomaterials Engineering
  • Protein Engineering

Background:

  • Self-organization is crucial for complex biological structures but challenging to engineer in synthetic systems.
  • Intrinsically disordered proteins (IDPs) lack stable structures and are key to cellular self-organization.
  • Engineering controllable multicellular materials requires precise control over cell-cell interactions.

Purpose of the Study:

  • To present a generalizable platform for high-density display of IDPs on Escherichia coli.
  • To engineer programmable, self-organizing living materials using IDPs.
  • To demonstrate dynamic control over cellular condensate formation and macroscopic material properties.

Main Methods:

  • Developed the intrinsically disordered protein display platform (iDP^2) using CsgF as a scaffold for surface display on E. coli.
  • Fused and presented IDP domains lacking stable tertiary structure on the bacterial surface.
  • Programmed cells with orthogonal IDPs to achieve sequence-specific segregation and form dynamic cellular condensates.
  • Investigated environmental cue (ionic strength, temperature) influence on condensate aggregation state.
  • Processed condensates via extrusion to generate macroscale filaments.

Main Results:

  • Successfully displayed IDPs at high density on E. coli, favoring disordered sequences.
  • Demonstrated that IDPs with phase separation propensity drive dynamic cellular condensate formation.
  • Achieved sequence-specific segregation of mixed cell populations using orthogonal IDPs, creating spatially organized living assemblies.
  • Showcased dynamic tunability of condensate aggregation state by environmental cues, predictable from IDP phase behavior.
  • Generated macroscale filaments from condensates that maintained structural integrity and population segregation.

Conclusions:

  • The iDP^2 platform provides a programmable framework for controlling cell-cell interactions and engineering living materials.
  • This approach enables the design of adaptive, self-organizing biological systems with customizable properties.
  • Disordered protein motifs are versatile tools for creating responsive and multicellular engineered biological systems.