Figure 1
From: A CRISPR-engineered isogenic model of the 22q11.2 A-B syndromic deletion
Proof-of-principle CRISPR engineering of 22q11.2 A-D deletion in HEK293T cells. (A) Cytogenetic representation of the 22q11.2 locus on chromosome 22 in red. The 22q11.2 A-D region within the locus is depicted by dashed lines. The low copy repeats (LCRs) (A), (B), (C) and (D) are marked by green boxes and key genes within the region are marked. The representative breakpoints resulting from gRNA1 targeting the LCRs A and D flanking the ~ 3 Mb deletion, and gRNA2 targeting LCRs A and B flanking the ~ 1.5 Mb deletion, are represented by purple and pink arrows, respectively. (B) qPCR screening of single-cell clones after genome editing with gRNA1, showing reduction in copy number of genes (TBX1, COMT, CRKL, HIRA) within the A-D deletion region in comparison to genes (MAPK1 and BID) flanking the deleted region. Fold-change of genomic qPCR signal shown in comparison to parental, unmodified HEK293T cells. n = 3 technical replicates, + /– S.D. shown. (C) Representative SNP microarray data from a non-edited control (top) and 22q11.2 deletion HEK293T clone (bottom) confirms deletion of the A-D region. (D) Shotgun proteomics reveals a decrease in expression of proteins encoded within the 22q11.2 A-D region (in red) compared to other quantified proteins encoding on flanking regions on chromosome 22 (in black). Data shows mean-log2 fold change per protein from n = 3 deletion and n = 3 control (non-edited) HEK293T clones, each expanded from single-cell selections.
