Fig. 4From: Universal CARs, universal T cells, and universal CAR T cellsProduction of universal TCR−/HLA− T cells using ZFN (zinc finger nuclease). a The T cells were obtained from healthy adult donors. ZFN mRNA pairs were delivered to the T cells by electroporation. ZFN pairs designed to target TCR αor βconstant regions and HLA-A lead to DNA double-strand breaks at the given sites and cause deletion or insertion of nucleotides, resulting in permanent loss of gene expression. These TCR−/HLA− T cells made from allogeneic healthy donors can be used as universal T cells for preparation of “off-the-shelf” CAR T cells. b Structure of the ZFN pair. One ZFN pair was designed to bind exon 1 of the TCRα constant region (TRAC), and the underlined nucleotide sequences represent the targeted binding sequences. The red blocks represent coding regions and the black block represents a noncoding region (adapted from Torikai H, et al. Blood 2012; 119(24):5697–5705)Back to article page