Inflammation conditional genome editing mediated by the CRISPR-Cas9 system.

Area of Science:

• Molecular Biology
• Gene Editing
• Immunology

Background:

• CRISPR-Cas9 specificity in pathological conditions is not well understood.
• Inflammation triggers cellular responses involving signaling pathways like NF-κB.

Purpose of the Study:

• To engineer an inflammation-inducible CRISPR-Cas9 system (NBS-CRISPR).
• To investigate the therapeutic potential of targeting inflammatory genes.
• To develop a system minimizing off-target effects during inflammation.

Main Methods:

• Grafting an NF-κB binding sequence to CRISPR-Cas9 to create NBS-CRISPR.
• Targeting the MyD88 gene to reverse inflammatory conditions.
• Constructing an NBS-P65-CRISPR system to fuse Cas9 with P65 to prevent leaky activity.

Main Results:

• NBS-CRISPR's genetic scissor function is activated by inflammation.
• Therapeutic effects were achieved by targeting the MyD88 gene.
• The NBS-P65-CRISPR system reduced unwanted Cas9 activity.

Conclusions:

• Inflammation-inducible CRISPR-Cas9 offers a novel strategy for pathological gene interrogation.
• This approach provides new avenues for studying and treating inflammatory diseases.
• The developed system enhances specificity and therapeutic potential in genome editing for inflammation.