The clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated proteins (Cas) system is a bacterial defense mechanism that uses RNA-guided enzymes to cut specific sequences of DNA or RNA. This property of the CRISPR-Cas system can be harnessed for diagnostic purposes by designing guide RNAs (gRNA) that target specific sequences. Recently, CRISPR-based diagnostics approaches have been intensively developed due to sensitivity, selectivity, and versatility. They are already in use for detecting mutations associated with cancer and inherited diseases and are being developed for a wide range of other applications, such as viral infections and inherited disorders. Herein, we introduce the CRISPR-based diagnostic methods recently developed in KRIBB. By using various kinds of Cas proteins, the methods allow for highly specific and sensitive detection of disease-causing mutations in a simple and cost-effective manner. The platforms can be adapted for the detection of a wide range of mutations and have the potential to be valuable tools for both research and clinical applications.

Learning Objectives:

1. Describe how CRISPR/Cas systems can be adapted for diagnostic applications.

2. Differentiate between various Cas proteins used in CRISPR-based diagnostic platforms.

3. Explain how CRISPR-based diagnostics achieve sensitivity and specificity in detecting disease-causing mutations.