Next-generation CRISPR/Cas-based ultrasensitive diagnostic tools: current progress and prospects.

Area of Science:

• Molecular Biology
• Biotechnology
• Genetics

Background:

• CRISPR/Cas systems, including SpCas9, are established gene editing tools.
• Cas12 and Cas13 variants exhibit unique trans-cleavage activity for nucleic acid recognition.
• CRISPR diagnostics utilize Cas12/13 for sensitive detection via fluorescent signals.

Purpose of the Study:

• To review the application of CRISPR/Cas systems as next-generation biosensors.
• To consolidate experimental evidence on CRISPR-based diagnostic technologies.
• To highlight the role of Cas12 and Cas13 trans-cleavage in biosensing.

Main Methods:

• CRISPR/Cas gene editing mechanisms (NHEJ, HDR).
• Cas12/Cas13 effector variants and their trans-cleavage activity.
• CRISPR diagnostics utilizing Ribonucleoproteins (RNPs) and fluorescent probes.
• Integration with nanotechnology and mobile-based detection.

Main Results:

• Cas12/13 mediated trans-cleavage enables sensitive target nucleic acid recognition.
• CRISPR diagnostics generate quantifiable fluorescent signals upon target binding.
• Nanotechnology integration enhances feasibility and enables lateral flow-based detection.

Conclusions:

• CRISPR/Cas systems, especially Cas12 and Cas13, show significant potential as next-generation biosensors.
• The trans-cleavage activity is crucial for the high sensitivity of CRISPR-based diagnostics.
• CRISPR diagnostics are becoming more feasible with advancements in nanotechnology and mobile integration.