CRISPR/Cas12a: A Comprehensive Review from Structural Foundations to Applications in Nucleic Acid Precision Detection

Area of Science:

• Biotechnology
• Molecular Biology
• Biosensing

Background:

• CRISPR/Cas12a, known for gene editing, possesses unique trans-cleavage activity.
• This activity enables its evolution into a versatile molecular detection tool.

Purpose of the Study:

• To systematically review the structural basis and molecular mechanism of CRISPR/Cas12a for molecular detection.
• To highlight its signal amplification capabilities and diverse applications.
• To discuss current challenges and future prospects in biosensing.

Main Methods:

• Systematic review of CRISPR/Cas12a technology.
• Analysis of its mechanism for converting nucleic acid recognition into signal amplification.
• Exploration of integration with amplification and detection platforms.

Main Results:

• CRISPR/Cas12a effectively converts specific nucleic acid recognition into cascade signal amplification.
• Applications include pathogen diagnosis, species identification, food safety, and traditional Chinese medicine authentication.
• Integration with isothermal amplification and multimodal platforms enhances portability, visualization, and quantification.

Conclusions:

• CRISPR/Cas12a is a powerful platform for molecular diagnostics and biosensing.
• Further development requires addressing sensitivity, accuracy, crRNA design, and standardization.
• Convergence with microfluidics and AI promises next-generation precision biosensing.