A highly efficient transgene knock-in technology in clinically relevant cell types
Inefficient knock-in of transgene cargos limits the potential of cell-based medicines. In this study, we used a CRISPR nuclease that targets a site within an exon of an essential gene and designed a cargo template so that correct knock-in would retain essential gene function while also integrating the transgene of interest. Cells with non-productive insertions and deletions would undergo negative selection.
, achieved knock-in efficiencies of more than 90% in clinically relevant cell types without impacting long-term viability or expansion. knock-in rates in T cells are more efficient than state-of-the-artknock-in with AAV6 and surpass more than 90% efficiency even with non-viral DNA cargos. As a clinical application, natural killer cells generated from induced pluripotent stem cells containing
knock-in of CD16 and mbIL-15 show substantially improved tumor killing and persistence in vivo.
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