CRISPR/Cas‐SERS Sensing Platforms: A Frontier Technology for Next‐Generation Fast, Low‐Cost, Ultra‐Micro Biosample Detection

ABSTRACT Researchers have long been interested in nucleic acid detection technology. Surface‐enhanced Raman spectroscopy (SERS) is distinguished by its high sensitivity, minimal sample volume, resistance to fluorescence interference, cost‐effectiveness, and rapidity compared to conventional nucleic acid analysis methods. The clustered regularly interspaced short palindromic repeats/CRISPR‐associated protein (CRISPR/Cas), a novel gene editing tool, has garnered significant interest in nucleic acid analysis due to its precise identification and isothermal advantages. Integrating the CRISPR/Cas system's specific identification capabilities with the high‐sensitivity fingerprinting properties of SERS offers a sensitive, ultra‐low volume, rapid, and straightforward method for detecting new nucleic acid modalities. This review delineates the components and characteristics of the CRISPR/Cas system, encompassing three Cas proteins (Cas9, Cas12, and Cas13) and detection technologies derived from CRISPR/Cas, namely, high‐sensitivity enzymatic reporter unlocking (SHERLOCK) and DNA endonuclease‐targeted CRISPR trans reporters (DETECTR). Advancements in SERS and CRISPR/Cas‐SERS‐based nucleic acid assays were emphasized, encompassing traditional SERS and CRISPR/Cas‐SERS‐based nucleic acid and non‐nucleic acid tests. Examples encompass microfluidics/microdroplet‐based CRISPR/Cas‐SERS and non‐amplified detection based on CRISPR/Cas‐SERS, and so on.