From cellular therapy to biologics: Non-viral delivery strategies for in vivo CAR-T engineering

Although CAR-T cell therapy has achieved breakthrough progress in the treatment of hematological malignancies, its current reliance on ex vivo manufacturing presents significant limitations in terms of efficiency, cost, and potential impact on cellular functionality. In vivo CAR-T strategies are increasingly recognized as a pivotal pathway toward the scalable and widely applicable deployment of CAR-T technologies. Among these, non-viral delivery systems have emerged as one of the principal enabling approaches. This review provides an overview of the key physiological barriers encountered in in vivo CAR-T cell engineering, alongside the corresponding principles of engineered design. It further outlines the evolving trends in the chemical composition of delivery materials and discusses the functional roles of various genetic payloads within in vivo CAR-T therapy. The objective of this comprehensive survey is to provide a forward-looking perspective to guide the rational design and clinical translation of non-viral-mediated in vivo CAR-T technologies.