Owing to the advent of the chimeric antigen receptor (CAR), cell-based immunotherapy has provided a unique avenue to treating oncological malignancies that are currently limited by traditional treatments such as chemotherapy and radiation. More recently, the use of engineered natural killer (NK) cells expressing CARs (CAR-NK) have been shown to possess an improved safety profile compared to CAR-T cells, making CAR-NKs a desirable treatment option for many cancers. However, primary sources for NK cells (peripheral blood and umbilical cord blood) present challenges such as the low yield of NK cells and donor heterogeneity making it difficult to generate standardized treatments. Nonetheless, CAR-NK cells have shown to be efficacious in treating various leukemic and lymphoid cancers amenable to allogeneic “off-the-shelf” based therapies.  To advance the development of these off-the-shelf therapies, we have leveraged genome engineering approaches to advance human induced pluripotent stem cells as cell chassis towards the development of CAR-NK therapies. Towards this goal, we have evaluated different iPSC reprogramming and genome targeting approaches to generate iPSCs and introduce CAR in an effort to improve CAR expression.  We validated a novel serum-free defined differentiation strategy to generate a pure population of CD56+ CD3-NK (iNK) cells. The iNK cells expressed a robust panel of requisite NK specific activating and inhibitory receptors as well as a exhibiting a robust cytotoxicity profile in response to target cancer cells. Collectively, we have optimized genome engineering approaches and a defined serum free differentiation and biomanufacturing approach to generated iPSC-derived NK cells that can be leveraged towards developing novel CAR-based immunotherapies for improved accessibility and affordability of iPSC-derived immunotherapy cell products.