Metal-Organic Framework as a Bioorthogonal Catalyst for Gene Editing

Area of Science:

• Materials Science
• Chemical Biology
• Biotechnology

Background:

• Metal-organic frameworks (MOFs) offer tunable structures for various applications.
• Bioorthogonal chemistry enables reactions within living systems without interference.
• RNA manipulation is crucial for gene editing and disease treatment.

Purpose of the Study:

• To design MOFs with molecularly defined catalytic pores for bioorthogonal RNA manipulation.
• To investigate the influence of MOF structural precision on RNA recognition and conversion.
• To develop MOF-based catalysts for intracellular RNA reactions and therapeutic applications.

Main Methods:

• Reticular design and synthesis of MOF libraries with varied Cu cluster geometries and pore dimensions.
• Small-angle X-ray scattering (SAXS) to study substrate positioning within MOF pores.
• Synchrotron soft X-ray microscopy for visualizing intracellular MOF localization and integrity.
• In vitro and in vivo experiments for RNA cleavage, formation, and gene editing regulation.

Main Results:

• MOFs with dimensionally matched mesopores and Cu(I) clusters efficiently catalyzed RNA bond cleavage and formation in living systems.
• Pore chemistry was found to govern substrate positioning, enhancing RNA engagement and product release for complete conversion.
• Intracellular visualization confirmed MOF integrity and bioorthogonal catalytic activity.
• An amine-functionalized MOF (MOF-248-NH2) demonstrated controlled sgRNA activation for CRISPR/Cas9 gene editing and cardiac disease treatment in a canine model.

Conclusions:

• Reticular pore engineering is a powerful strategy for creating programmable bioorthogonal MOF catalysts.
• MOFs can be designed for precise control over RNA manipulation within living cells.
• This approach holds significant potential for advancing gene editing technologies and developing novel therapeutics.