Application of extracellular vesicles in the CRISPR-based diagnosis and treatment: possibilities and challenges

The CRISPR–Cas system has revolutionized molecular diagnostics and gene editing, yet its clinical translation is hindered by delivery barriers, off-target activity, immunogenicity, and manufacturing challenges. Compared with viral vectors and synthetic non-viral carriers such as lipid nanoparticles, extracellular vesicles (EVs) offer a biologically derived delivery platform with superior biocompatibility, reduced immunogenicity, intrinsic cargo protection, and natural barrier-crossing capability. Engineered EVs can further achieve cell- or tissue-specific targeting. In diagnostics, endogenous EV proteins and nucleic acids provide disease-informative signatures that can interface with CRISPR nuclease readouts for highly sensitive detection. This review summarizes the therapeutic and diagnostic potential of EV-CRISPR platforms, covering strategies for loading CRISPR cargos (producer-cell engineering, post-isolation methods), cargo formats, and surface targeting approaches. We evaluate preclinical performance with attention to biodistribution, safety, innate and adaptive immune responses, and genomic integrity, as well as analytical assays and scalable manufacturing considerations essential for clinical translation. Finally, we discuss emerging opportunities, including AI-guided optimization of EV-CRISPR design and integrated EV platforms that combine disease detection with therapeutic intervention, highlighting their promise for advancing precision medicine.