CRISPR/Cas-based nucleic acid detection strategies: Trends and challenges

Area of Science:

• Molecular Biology
• Biotechnology
• Biosensor Technology

Background:

• CRISPR/Cas systems are versatile tools for nucleic acid detection and biosensing.
• Diverse CRISPR/Cas systems (e.g., Cas9, Cas12, Cas13) employ distinct mechanisms for sequence and activity detection.
• These systems are frequently integrated with amplification techniques like PCR and LAMP for enhanced diagnostics.

Purpose of the Study:

• To provide a comprehensive overview of CRISPR/Cas-based nucleic acid detection methods.
• To highlight recent advancements in CRISPR/Cas detection system development and components.
• To discuss the operational challenges, advantages, and disadvantages of various CRISPR/Cas systems.

Main Methods:

• Review of existing literature on CRISPR/Cas systems for nucleic acid detection.
• Analysis of various CRISPR/Cas systems, including Cas9, Cas12, Cas13, Cas14, and Cas3.
• Examination of integration strategies with amplification technologies (PCR, LAMP, RPA).

Main Results:

• CRISPR/Cas-based biosensors are effective for detecting viral and bacterial pathogens in clinical samples.
• Applications extend to biosecurity, food safety, and environmental monitoring.
• CRISPR/Cas systems demonstrate superior specificity compared to traditional molecular diagnostic methods.

Conclusions:

• CRISPR/Cas technology offers significant potential for sensitive and specific nucleic acid detection.
• Further research is needed to address operational challenges and optimize system components.
• Considerations for improving CRISPR/Cas-based nucleic acid testing are crucial for future development.