Figure 1.

Generation and evaluation of simultaneous knockout of the HLA-A, HLA-B, and CIITA genes in iPSCs by CRISPR-Cas9

(A) The design of a gRNA (HLA-A24-ex2g1) to simultaneously target the HLA-A and HLA-B genes, and another gRNA (CIITA-ex3g5) to target the CIITA gene. The genomic coordinate is based on hg19. (B) Histograms of the immunoreactivity intensity of the anti-HLA-A and anti-HLA-B antibodies for bulk genome-edited iPSCs. (C) The workflow of the production of genome-edited iPSC clones and their evaluation. (D) Sanger sequencing of the genome-edited iPSCs after single-cell cloning. Insertions or deletions at the target sites in the genome-edited iPSC clones were confirmed by electropherograms. (E) Size distribution of insertions or deletions at each target gene locus.