CRISPR in Immuno-Oncology: Engineering CAR-T and Next-Generation Immune Cells

Abstract

Immuno-oncology has reshaped the therapeutic landscape of cancer treatment by shifting focus from directly targeting tumor cells to mobilizing the immune system against malignancies. Among the most transformative advances in this field is the development of chimeric antigen receptor T-cell therapy, which has demonstrated remarkable efficacy in hematologic cancers. However, persistent challenges such as limited durability, immune escape, toxicity, and poor performance in solid tumors have constrained its broader clinical impact. The emergence of clustered regularly interspaced short palindromic repeats (CRISPR) genome editing has introduced a powerful and versatile platform for engineering immune cells with enhanced specificity, persistence, and functionality. CRISPR-based approaches enable precise gene knockout, targeted gene insertion, epigenetic modulation, and multiplex editing, allowing researchers to redesign immune cells at multiple regulatory levels. These capabilities have significantly advanced CAR-T cell engineering and have catalyzed the development of next-generation immune effectors, including natural killer cells, macrophages, and stem cell-derived immune populations. Furthermore, CRISPR technology has opened new avenues for overcoming the immunosuppressive tumor microenvironment, improving safety profiles, and enabling scalable, off-the-shelf therapies. This review provides a comprehensive examination of CRISPR applications in immuno-oncology, with an emphasis on CAR-T optimization and the engineering of next-generation immune cells. It discusses mechanistic foundations, technological innovations, preclinical and clinical advancements, safety considerations, and future directions. Collectively, CRISPR-driven immune engineering represents a paradigm shift toward more precise, effective, and accessible cancer immunotherapies.