Programmable RNA editing by recruiting endogenous ADAR using engineered RNAs

Area of Science:

• Molecular Biology
• Genetic Engineering
• RNA Therapeutics

Background:

• Current RNA editing tools face challenges like immunogenicity and delivery barriers.
• Exogenous proteins or modified guide RNAs can cause unwanted effects.

Purpose of the Study:

• To introduce a novel, efficient, and specific RNA editing system.
• To overcome limitations of existing RNA editing technologies.

Main Methods:

• Developed LEAPER (leveraging endogenous ADAR for programmable editing of RNA) using engineered ADAR-recruiting RNAs (arRNAs).
• Delivered arRNAs via plasmids, viral vectors, or synthetic oligonucleotides.
• Tested editing efficiency, specificity, and cellular activity in various cell types.

Main Results:

• Achieved up to 80% RNA editing efficiency with LEAPER.
• Demonstrated high specificity with rare off-target edits.
• Showed LEAPER's activity across diverse cell types, including primary human cells.
• Restored enzyme activity in Hurler syndrome fibroblasts without immune response.

Conclusions:

• LEAPER offers a precise and efficient single-molecule system for RNA editing.
• The approach has broad applicability for therapeutic interventions and basic research.
• LEAPER minimizes immunogenicity and delivery challenges associated with other methods.