AAV capsid evolution in human T cells identifies a family of CD7-targeting variants for efficient T and NK cell engineering [CRISPR screen]

Structural analyses revealed that the APR substitution induced a spatial conformation change in the capsid surface, repositioning the VR-IV domain to improve accessibility and cooperative binding with CD7. We further demonstrate that AAV.APR31 enables robust transgene delivery, supports high-efficiency CRISPR-Cas9-mediated knock-in of large constructs at multiple genomic loci, and facilitates AAV-transposon-mediated DNA integration.

Notably, AAV.APR31 has been successfully applied to generate a variety of therapeutic immune cell products, including conventional CAR-T cells, bispecific CAR-T cells, synthetic TCR-T cells, and CAR-NK cells. The development of these synthetic capsids expands the toolbox for precise genome engineering and holds strong translational potential for both basic research and next-generation human T and NK cell-based cellular immunotherapies.