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Updated: Jun 26, 2025

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Expanding RNA editing toolkit using an IDR-based strategy.

Minghui Di1,2, Junjun Lv1,2, Zhengyu Jing1,2

  • 1School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.

Molecular Therapy. Nucleic Acids
|May 9, 2024
PubMed
Summary
This summary is machine-generated.

Researchers developed a new human-origin RNA base editor for C to U editing. Fusing a histidine-rich domain enhanced its efficiency, demonstrating a general method to improve RNA base editors.

Keywords:
CIRTSIDRLLPSMT: RNA/DNA EditingRNA base editingcompacthuman originimmunogenicity

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetic Engineering

Background:

  • RNA base editing offers precise genetic modification but faces challenges in efficiency and specificity for C to U changes.
  • Existing editors often lack ideal characteristics such as being compact, efficient, and non-immunogenic.

Purpose of the Study:

  • To develop a compact, fully human C to U RNA base editor.
  • To enhance the efficiency of RNA base editors using a novel strategy.

Main Methods:

  • Fusion of the human C to U editing enzyme RESCUE-S with the Cas inspired RNA targeting system (CIRTS) to create CIRTS-RESCUEv1 (V1).
  • Incorporation of a histidine-rich domain (HRD) from human CYCT1, known for liquid-liquid phase separation (LLPS), into the V1 editor.
  • Evaluation of editing efficiency and mechanism, including LLPS-dependent puncta formation.

Main Results:

  • The initial V1 editor showed low efficiency.
  • Fusion with HRD significantly enhanced on-target editing efficiency of V1.
  • HRD-mediated enhancement was linked to liquid-liquid phase separation (LLPS), with other intrinsically disordered regions (IDRs) also showing potentiation.
  • The HRD fusion strategy proved applicable to other C to U RNA editors.

Conclusions:

  • A novel, compact, human-origin C to U RNA base editor (CIRTS-RESCUEv1) was developed.
  • Incorporation of intrinsically disordered regions, leveraging liquid-liquid phase separation, is a general and effective strategy to enhance C to U RNA base editor efficiency.
  • This work expands the toolkit for RNA editing and provides a method to improve editor performance.