Rational Engineering of a Modular Group I Ribozyme to Control Its Activity by Self-Dimerization

Area of Science:

• Molecular Biology
• RNA Therapeutics
• Nanotechnology

Background:

• Group I ribozymes are catalytic RNA molecules with diverse functions.
• Current RNA nanotechnology often relies on small, limited functional motifs.
• Controlling ribozyme activity through self-assembly is an emerging area.

Purpose of the Study:

• To design and construct a dimer of the Tetrahymena group I ribozyme regulated by self-dimerization.
• To demonstrate the feasibility of using large RNA motifs for constructing functional RNA structures.
• To advance RNA nanotechnology by overcoming limitations of small functional units.

Main Methods:

• Rational design of a ribozyme dimer using P5abc and ΔP5abc domains.
• Modular engineering of RNA interaction motifs for programmed self-dimerization.
• Electrophoresis mobility shift assay (EMSA) to confirm dimerization.
• Ribozyme activity assays to validate catalytic function and regulation.

Main Results:

• Successful construction of a large, functional ribozyme dimer (tecto-GIRz).
• Demonstration that self-dimerization can regulate ribozyme activity.
• Validation of the strategy for incorporating large RNA motifs into functional structures.

Conclusions:

• The engineered ribozyme dimer expands the scope of RNA nanotechnology.
• Modular design principles allow for programmed self-assembly and regulation of large RNA structures.
• This work provides a foundation for developing complex, functional RNA-based nanomaterials.