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Updated: Sep 4, 2025

Large-scale Production of Recombinant RNAs on a Circular Scaffold Using a Viroid-derived System in Escherichia coli
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Engineering circular RNA for enhanced protein production.

Robert Chen1, Sean K Wang1,2, Julia A Belk3

  • 1Center for Personal Dynamic Regulomes, Stanford University, Stanford, CA, USA.

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Summary
This summary is machine-generated.

Synthetic circular RNAs (circRNAs) can be engineered for protein production. This study optimized circRNA design elements, significantly boosting protein yields for gene expression applications.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Gene Expression

Background:

  • Circular RNAs (circRNAs) are recognized for their stability and prevalence in eukaryotic cells, originating from back-splicing.
  • Both synthetic and endogenous circRNAs have demonstrated the capacity to encode proteins, highlighting their potential as a gene expression platform.

Purpose of the Study:

  • To develop a systematic method for the rapid assembly and evaluation of features influencing protein production from synthetic circRNAs.
  • To optimize key design elements for maximizing protein translation from synthetic circRNAs.

Main Methods:

  • Systematic optimization of five critical elements: vector topology, 5' and 3' untranslated regions, internal ribosome entry sites, and synthetic aptamers.
  • Testing and validation of optimized design principles for enhanced circRNA translation.

Main Results:

  • Achieved several hundred-fold improvement in circRNA protein yields through optimized design.
  • Demonstrated superior and more durable protein translation from optimized circRNAs compared to messenger RNA (mRNA) in vitro and in vivo.
  • Confirmed the generalizability of these design principles across various transgenes.

Conclusions:

  • The developed design principles significantly enhance protein production from synthetic circRNAs.
  • Optimized synthetic circRNAs offer a promising and efficient platform for gene expression with improved translation efficiency and durability.
  • This systematic approach provides a robust strategy for engineering circRNAs for biotechnological applications.