Amplification-Free Molecular Diagnostics with Real-Time and Spectral Multiplexing Capabilities Using CRISPR-Mediated SERS

• Applicant: Namhyun Choi, Ph.D.
• Subject Area: Analytical Chemistry
• Term: since 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 550274647

The groundbreaking discovery of CRISPR/Cas technology has attracted significant attention in many areas of biomedicine due to its immense potential. It has also been applied in molecular diagnostics for detecting target DNA/RNA. However, time-consuming pre-amplification steps currently limit the widespread use of this technology, particularly for time-critical high-risk diseases. Therefore, the aim of this project is to establish an amplification-free analytical platform for rapid and highly sensitive simultaneous detection of multiple target oligonucleotides for comprehensive diagnostics. This goal can be achieved by using surface-enhanced Raman scattering (SERS) as an optical detection technique. Establishing the multiplexed real-time detection of multiple oligonucleotides through CRISPR-mediated SERS (SERS-CRISPR) requires a multidisciplinary approach covering the following areas: colloid chemistry (uniform, photostable, and bright SERS nanotags), analytical chemistry (optical spectroscopy), biophysical chemistry (enzyme kinetics), and engineering (module development and system integration). The envisioned SERS-CRISPR platform for molecular diagnostics is expected to be capable of real-time detection of a panel of multiple oligonucleotides in less than 20 minutes with attomolar sensitivity, which is significantly faster than PCR as the current gold standard. This will pay the way for next generation on-site molecular diagnostics.

• DFG Programme: Research Grants