Figure 2
From: A CRISPR-engineered isogenic model of the 22q11.2 A-B syndromic deletion
CRISPR engineering of 22q11.2A-B deletion in iPS cells. (A) Schematic of strategy for CRISPR engineering of 22q11.2A-B deletion in iPS cells containing inducible Cas9 by using the SCORE approach, followed by differentiation into neurons and functional analyses. (B) Cytogenetic representation of the chromosome 22q11.2 locus including RefSeq-annotated protein-coding and non-coding genes. Image adapted from UCSC Genome Browser (Human GRCh38/hg38). (C) Single cell sorting and clonal expansion of iPS cells yielded 68 total clones expanding for > 2 weeks post-sort, 5 of which screened positive by qPCR assay for the 22q11.2 A-B deletion, 3 of which were confirmed by genome-wide SNP array to harbor the 22q11.2A-B deletion and no other large genomic alterations (see Supplementary Table 1) and were used for further study. (D) Representative SNP microarray data from a 22q11.2A-B deletion clone (bottom) with comparison to control clone (top). (E) Representative images of characterization of CRISPRn/WTC-11 control and 22q11.2A-B deletion iPSC clones for stem cell markers OCT4 (magenta), SSEA4 (green) and SOX2 (red). Nuclei are labeled with Hoechst (blue). Scale bar = 400 μm. (F) Representative images of characterization of control and 22q11.2 A-B deletion NPC clones for expression of NPC marker nestin (green). Nuclei are labeled with Hoechst (blue). 40 × magnification. Scale bar = 100 μm.
