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Synthetic multistability in mammalian cells.

Ronghui Zhu1, Jesus M Del Rio-Salgado1, Jordi Garcia-Ojalvo2

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Researchers engineered a synthetic gene circuit called MultiFate for controllable multistability in mammalian cells. This circuit enables long-term, expandable cell state control, advancing synthetic biology and cell fate engineering.

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Area of Science:

  • Synthetic biology
  • Gene regulatory networks
  • Mammalian cell engineering

Background:

  • Multicellular organisms utilize gene regulatory circuits for cell fate determination.
  • Multistability, the generation of heritable cell states, is crucial for development.
  • Engineering synthetic multistable circuits is key to understanding and controlling cell fate.

Purpose of the Study:

  • To design and create a synthetic gene circuit for controllable and expandable multistability in mammalian cells.
  • To investigate the potential of engineered transcription factors in generating multiple stable cell states.
  • To lay the groundwork for engineering complex multicellular behaviors.

Main Methods:

  • Developed MultiFate, a synthetic circuit inspired by natural gene regulatory mechanisms.
  • Utilized engineered zinc finger transcription factors that self-activate and mutually inhibit.
  • Employed model-based design to engineer circuits capable of generating multiple stable states.
  • Validated long-term stability and controllability of engineered cell states.

Main Results:

  • Successfully engineered MultiFate circuits capable of generating up to seven distinct, mitotically heritable cell states.
  • Demonstrated long-term stability of these states for at least 18 days.
  • Showcased controlled state switching and modulation of state stability via external inputs.
  • Confirmed the expandability of the MultiFate circuit with additional transcription factors.

Conclusions:

  • MultiFate provides a robust platform for achieving long-term, controllable, and expandable multistability in mammalian cells.
  • This synthetic circuit offers insights into natural cell fate control mechanisms.
  • The findings establish a foundation for engineering sophisticated multicellular programs and behaviors.