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Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
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Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
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Enhancing Circular RNA Translation Efficiency Through Dual Internal Ribosome Entry Sites.

Yawen Sun1, Yimin Zhang2, Weijie Chen2

  • 1College of Biological Science and Medical Engineering, Donghua University, Shanghai 201620, China.

Biology
|February 26, 2026
PubMed
Summary
This summary is machine-generated.

Circular RNAs (circRNAs) offer stable drug delivery. A dual-internal ribosome entry site (IRES) strategy enhances circRNA translation efficiency, overcoming limitations of cap-independent expression for therapeutic applications.

Keywords:
EMCVIREScircRNAtranslation efficiency

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Drug Delivery Systems

Background:

  • Circular RNA (circRNA) is a stable, non-coding RNA molecule with potential as a drug delivery vector.
  • circRNAs lack a 5' cap, relying on internal ribosome entry sites (IRES) for translation initiation, which is less efficient than cap-dependent translation.
  • Enhancing circRNA translation is crucial for maximizing its therapeutic efficacy.

Purpose of the Study:

  • To develop a strategy for enhancing circRNA translation efficiency.
  • To investigate the efficacy of a dual-IRES system for cap-independent translation of circRNAs.
  • To identify optimal IRES elements for improved circRNA expression.

Main Methods:

  • Designed and constructed circRNA molecules incorporating a dual-IRES strategy.
  • Tested various IRES elements, focusing on those from the Encephalomyocarditis virus (EMCV) family.
  • Evaluated the translational activity and expression levels of engineered circRNAs with different IRES combinations.

Main Results:

  • A dual-IRES strategy significantly enhanced circRNA translation compared to single IRES elements.
  • EMCV-derived IRES elements, particularly when placed at the 3' of the coding sequence (CDS) in coordination with a 5' IRES, showed optimal enhancement.
  • The identified dual-IRES combinations were compatible with multiple coding sequences.

Conclusions:

  • The dual-IRES strategy is an effective method for boosting circRNA translation.
  • This approach overcomes the inherent limitations of cap-independent translation in circRNAs.
  • The findings provide a valuable tool for developing enhanced circRNA-based therapeutics and delivery systems.