Figure 2. Generation of ICAM-1 Knock-out Pluripotent Stem Cell (PSC) Lines.

ICAM-1 was knocked-out (KO) via CRISPR/Cas9 editing. (A) Schematic demonstrating the CRISPR/Cas9 KO strategy. The wild type (WT) codon (CTG) in the first exon of the ICAM-1 sequence is edited into a stop codon (TGA) via the addition of a nucleotide. (B) Representative PSC colony morphology (10X). (C) Pluripotency marker SSEA-4 staining by flow cytometry. (D) Teratomas were grown in immune-deficient mouse hosts by intramuscular injection of the H9 ICAM-1 KO PSC line with Matrigel. Representative Hematoxylin and Eosin stained images of (top) gut [Endoderm], (middle) cartilage [Mesoderm], and (bottom) retinal pigment epithelium [Ectoderm] representing all three germ layers. Multiple cell lines were made in human embryonic stem cells (hESCs) and in human induced PSCs; data are representative of all lines.