Unmodificated stepless regulation of CRISPR/Cas12a multi-performance

Area of Science:

• Molecular Biology
• Biotechnology
• Nucleic Acid Nanotechnology

Background:

• CRISPR technology faces challenges in meeting diverse molecular biology application needs.
• Accurate control of CRISPR performance is crucial for advancing its capabilities.

Purpose of the Study:

• To propose a flexible CRISPR/Cas12a regulation strategy using nucleic acid nanotechnology.
• To enable fine-grained control over CRISPR-Cas12a activity, speed, specificity, and sensitivity.

Main Methods:

• Utilized nucleic acid nanotechnology to create external RNA accessories for CRISPR/Cas12a.
• Switched RNA accessories to modulate reaction kinetics and thermodynamics.
• Applied the strategy to regulate delayed activation of Cas12a for one-pot assays.

Main Results:

• Achieved fine and near-stepless regulation of CRISPR/Cas12a multi-performance.
• Significantly improved specificity, enhancing molecular detection accuracy and gene editing safety.
• Demonstrated delayed activation of Cas12a, overcoming compatibility issues in one-pot assays.

Conclusions:

• The proposed strategy offers pioneering flexibility and versatility for CRISPR/Cas12a regulation.
• This approach allows for precise control without modifying core CRISPR components.
• The strategy holds potential for unlocking deeper applications of CRISPR in specialized fields.