Supplementary Figure 2: Indel analysis of AAV–Cpf1-mediated multiplexed gene editing in human primary CD4+ T cells.
From: One-step generation of modular CAR-T cells with AAV–Cpf1
a, T7E1 endonuclease assay for genome editing of both PDCD1 and TRAC loci in bulk or TCR– sorted T cells using AAV–crTRAC;crPDCD1 array with AAV9 packaging. Arrowheads point to cleavage products. b, Targeted mutagenesis efficiency of both PDCD1 and TRAC loci estimated by deep sequencing. The top ten most frequent variants at TRAC and PDCD1 loci generated by AAV9–crTRAC;crPDCD1, with the associated variant frequencies in the boxes to the right. Red arrowheads indicate predicted cleavage sites. One representative sample’s data are shown from 3 infection replicates. c, T7E1 endonuclease assay of genome editing of both PDCD1 and TRAC loci in bulk or TCR– sorted T cells using AAV–crTRAC;crPDCD1 array with AAV6 packaging. Arrowheads point to cleavage products. d, Targeted mutagenesis efficiency of both PDCD1 and TRAC loci estimated by deep sequencing. The top ten most frequent variants at TRAC and PDCD1 loci generated by AAV6–crTRAC;crPDCD1, with the associated variant frequencies in the boxes to the right. Red arrowheads indicate predicted cleavage sites. One representative sample’s data are shown from 3 infection replicates.
