Tuning Covalent Organic Frameworks for Nucleic Acid Loading: Toward Creating a Hierarchical Artificial Cell

Area of Science:

• Synthetic Biology
• Materials Science
• Biotechnology

Background:

• Bottom-up artificial cell construction is crucial for life sciences and applications.
• Existing one-pot methods often yield oversimplified structures.
• Advanced hierarchical structures are needed for more complex artificial cell functions.

Purpose of the Study:

• To develop a novel strategy for constructing artificial cells with nucleus-like structures.
• To utilize covalent organic frameworks (COFs) for DNA encapsulation and compartmentalization.
• To integrate cell-free transcription and translation (CFTT) within these artificial nuclei.

Main Methods:

• Utilized covalent organic frameworks (COFs) with spatiotemporal strategies for linear and circular DNA encapsulation.
• Employed a template sacrifice method (COFcap-1) for encapsulating circular plasmids within COFs.
• Integrated cell-free transcription and translation (CFTT) systems within the COF structures.
• Constructed artificial nucleus logic gates and encapsulated COF-DNA complexes within liposomes.

Main Results:

• Successfully constructed nucleus structures using COFs for both linear and circular DNA.
• Demonstrated the crucial role of COF pore size and microenvironment on CFTT efficiency.
• Engineered three artificial nucleus logic gates, achieving information transmission between artificial organelles.
• Successfully encapsulated DNA-loaded COFs within liposomes to form hierarchical artificial cells.

Conclusions:

• This study presents a novel method for creating highly structured artificial cells using porous COFs.
• The developed COF-based nucleus structure enhances DNA encapsulation and CFTT efficiency.
• This work opens new avenues for constructing complex, hierarchical artificial cells for various applications.