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Engineered Allosteric RNA Editors Enable Compact, Stimulus-Responsive Post-Transcriptional Circuits.

Alexander M Marzilli1, John T Ngo1

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Researchers developed inducible Deaminases Acting on RNA (iDARs) for programmable control over synthetic messenger RNAs (mRNAs). These iDARs enable trigger-dependent protein translation or mRNA degradation, offering precise control over gene expression.

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Translational regulation is a key biological control mechanism.
  • Synthetic mRNAs require programmable control for advanced applications.

Purpose of the Study:

  • To introduce inducible Deaminases Acting on RNA (iDARs) for conditional RNA editing.
  • To develop a generalizable framework for controllable deaminases and post-transcriptional circuits.

Main Methods:

  • Designed autoinhibited deaminase domains (DDs) using a domain-insertion strategy.
  • Engineered iDARs responsive to small molecules, antigens, proteases, and light.
  • Created novel RNA substrates with stop codons for conditional translation or degradation.
  • Tuned IP6-binding pockets for precise regulation and dose-dependent control.

Main Results:

  • Developed various iDARs (chemiDARs, antiDARs, lysiDARs, optiDARs) with conditional RNA-editing activities.
  • Achieved dose-dependent readthrough translation with dynamic ranges over 100-fold at low-nanomolar drug concentrations.
  • Demonstrated "self-editing" polycistronic transcripts for trigger-dependent protein expression from delivered mRNAs.
  • Showcased iDARs' ability to link biochemical sensing to de novo translation or mRNA decay.

Conclusions:

  • iDARs provide a versatile platform for creating controllable RNA editors.
  • This technology enables the design of sophisticated post-transcriptional regulatory circuits.
  • iDARs facilitate precise control over synthetic mRNA function in cellular contexts.