CRISPR-based non-nucleic acid detection

Area of Science:

• Molecular Biology
• Biotechnology
• Diagnostics

Background:

• Clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) trans-cleavage activities are foundational to next-generation diagnostics.
• CRISPR systems are engineered to detect a wide range of non-nucleic acids (NNAs), including ions, small molecules, proteins, and bacteria.

Purpose of the Study:

• To review the principles and advantages of CRISPR-based non-nucleic acid (NNA) detection.
• To classify existing CRISPR-based NNA detection strategies.
• To discuss challenges and future directions for CRISPR-based NNA detection.

Main Methods:

• Classification of CRISPR-based NNA detection strategies into three categories: generation/depletion of free activators, crRNA synthesis, and active Cas effector reconstruction.
• Review of diverse strategies for NNA recognition and regulation of Cas trans-cleavage activities.

Main Results:

• CRISPR systems can be engineered for NNA detection using trans-cleavage reactions for signal output.
• Strategies involve specific NNA recognition and modulation of Cas trans-cleavage activity.
• Three main classes of strategies are identified for CRISPR-based NNA detection.

Conclusions:

• CRISPR-based NNA detection offers a versatile platform for various applications.
• Addressing current challenges can advance both clinical and nonclinical uses of CRISPR diagnostics.
• Further development is needed to optimize detection sensitivity, specificity, and applicability.