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A Ψ-Ψ codon-anticodon pairing in nonsense suppression and translational recoding.

Yi Pan1, Elzbieta Kierzek2, Ryszard Kierzek2

  • 1Department of Biochemistry and Biophysics, Center for RNA Biology, University of Rochester Medical Center, Rochester, NY, USA.

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|September 12, 2025
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Summary
This summary is machine-generated.

This study engineered guide RNAs to add pseudouridine (Ψ) to mRNA and tRNA, creating a novel Ψ-Ψ base pair. This unusual pair enhances codon recognition, promoting premature termination codon readthrough and translational recoding.

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Area of Science:

  • Molecular Biology
  • RNA Biology
  • Genetics

Background:

  • Pseudouridine (Ψ) is a modified uridine with unclear base-pairing flexibility.
  • Premature termination codons (PTCs) disrupt protein synthesis, leading to non-functional proteins.

Purpose of the Study:

  • To engineer artificial guide RNAs for targeted pseudouridylation of PTCs in mRNA and tRNA.
  • To investigate the impact of pseudouridylation on codon-anticodon recognition and PTC readthrough.

Main Methods:

  • Engineering artificial box H/ACA guide RNAs for targeted pseudouridylation.
  • Utilizing yeast and human cell systems to assess mRNA and tRNA modifications.
  • Employing melting curve analysis to determine the affinity of the modified base pair.

Main Results:

  • Targeted pseudouridylation created a novel Ψ-Ψ codon-anticodon pair.
  • The Ψ-Ψ pair significantly enhanced codon-anticodon recognition and PTC readthrough.
  • mRNA levels, tRNA stability, and tRNA charging remained unaffected by pseudouridylation.

Conclusions:

  • An unusual Ψ-Ψ base pair was identified, enhancing codon-anticodon recognition.
  • This modification robustly promotes PTC readthrough and translational recoding.
  • The findings offer a new strategy for modulating protein synthesis via RNA modification.