Detecting pathogens with Zinc-Finger, TALE and CRISPR- based programmable nucleic acid binding proteins.

Area of Science:

• Molecular Biology
• Biotechnology
• Genomics

Background:

• Engineered nucleic acid binding proteins, including Zinc-Finger, TALE, and CRISPR-based systems, have revolutionized genome editing.
• These programmable proteins enable highly specific DNA and RNA binding, paving the way for novel diagnostic platforms.

Purpose of the Study:

• To review the development and application of engineered nucleic acid binding proteins for pathogen detection.
• To highlight the potential of CRISPR-associated enzymes and associated diagnostic methods for sensitive and specific nucleic acid detection.

Main Methods:

• Leveraging engineered nucleic acid binding proteins like Zinc-Finger, TALE, and CRISPR systems.
• Utilizing novel CRISPR-associated enzymes such as Cas13a (C2c2), Cas12a (Cpf1), and Csm6.
• Developing diagnostic platforms like DNA Endonuclease-Targeted CRISPR Trans Reporter (DETECTR) and Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK).

Main Results:

• Demonstrated single-molecule resolution sensitivity for pathogen nucleic acid detection.
• Successfully detected nucleic acids from bacterial pathogens and viral threats like Zika virus.
• Showcased genotyping capabilities, differentiating between human papillomavirus (HPV) types 16 and 18.

Conclusions:

• Engineered nucleic acid binding proteins, particularly CRISPR-based systems, offer reliable and sensitive diagnostic capabilities.
• Advancements in multiplexing, quantification, and instrument-free detection will drive the adoption of these technologies for point-of-care diagnostics.
• These technologies hold significant potential for rapid and accurate detection of bacterial and viral infections.