CRISPR/dCAS9 genomic engineering for boosting the therapeutic potential of Extracellular Vesicles

This study aimed to enhance the therapeutic effect of MSC-derived extracellular vesicles (EVs) by overexpressing the TSG-6 gene using CRISPR activation and evaluate the biological activity of these modified EVs on human intervertebral disc (IVD) cells in vitro. n immortalized human MSC line was transduced with a CRISPR activation system targeting TSG-6. MSC-EVs were collected via ultracentrifugation, and their particle size/number was analyzed. Gene and protein expression were assessed to evaluate transduction efficiency. EV proteomics was analyzed by mass spectrometry. Human IVD cells were isolated from patients undergoing spinal surgery, pre-stimulated with IL-1β, and treated with MSC-EVs.