Abstract
Peripheral blood mononuclear cells (PBMCs) are harvested from patients and purified for CD19+ B cells. B cells can also be derived from iPSCs. Purified B cells are activated/expanded in culture to prime them for genome engineering. Lentivirus, transposons, and CRISPR/Cas9 together with recombinant adeno-associated viral vector (rAAV) carrying DNA donor template for homology directed repair (HDR) can be used to integrate a transgene into the B cell genome. CRISPR-based engineering tools can also be used for gene modification or knockout. Engineered B cells may be further expanded in culture.
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