Limb-girdle muscular dystrophy type 2A/R1 (LGMD2A/R1) is a rare form of muscular dystrophy with around 4000 patients in the US and approximately 80-100 new patients identified every year. LGMD2A/R1 is caused by mutations in the CAPN3 gene, which leads to loss of functional calpain-3 protein, resulting in symmetrical muscle wasting of the proximal limb and girdle muscles. There is currently no approved treatment for LGMD2A/R1. Regenerative therapies derived from induced pluripotent stem cells (iPSCs) may provide a promising approach to regenerate muscle mass associated with muscle wasting. Here we describe the pre-clinical development of an iPSC-based autologous therapy for LGMD2A/R1. A CRISPR-based knock-in strategy was developed to replace the endogenous defective CAPN3 gene with a functional version in LGMD2A/R1 patient-derived iPSCs. Using a chemically defined, step-wise myogenic lineage specification protocol, PAX7-expressing myogenic progenitor cells (MPCs) were then generated; upon myogenic fusion induction, these cells formed functional myotubes that produced CAPN3 mRNA and protein. We then transplanted CAPN3 edited LGMD2A/R1 iPSC derived MPCs into the injured TA muscles of NSG mice and demonstrated donor cell-derived muscle regeneration. Furthermore, a subpopulation of transplanted MPCs seeded in the local muscle stem cell niche area under the basal lamina in mice where they adopt a quiescent state and contribute to regeneration upon additional injury, thus providing evidence of a long-term maintenance and regenerative capability. To facilitate in vivo efficacy studies in an animal model of LGMD2A/R1, we generated an immunodeficient calpain-3 null mouse by back-crossing CAPN3KO mice (JAX: CC041-Capn3em10Lutzy/J) with NSG (JAX: NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) mice. Intramuscular transplantation of CAPN3 edited MPCs into injured muscle of CAPN3KO-NSG mice leads to donor-derived muscle fiber regeneration, supporting the rationale for continued development. Manufacturing of LGMD2A/R1 iPSC-derived MPCs to support IND-studies and first-in-human trials is ongoing with the goal of IND-filing.