Protocol for manipulating mRNA splicing via vivo-morpholino in early-activating muscle stem cells

Muscle stem cells (MuSCs), also called satellite cells (SCs), are essential for skeletal muscle regeneration. SCs undergo rapid splicing changes upon activation, hindering the study of alternative splicing during SC activation. Here, we describe a protocol for manipulating mRNA splicing in early-activating SCs during isolation using vivo-morpholino (Vivo-MO). We also detail steps for isolating vivo-morpholino-treated SCs, validating the splicing alteration by RT-PCR, and evaluating the functional outcome in SC activation using an EdU incorporation assay. For complete details on the use and execution of this protocol, please refer to Lin et al.1.