Figure 1.

Detection of chromosome 18q arm truncation following CRISPR/Cas9 targeting of ZNF516.

(A) Schematic of 7 exons of ZNF516 on the reverse strand, with coding sequence shown as filled rectangles, and untranslated regions as empty rectangles. crRNA target sites are indicated in green in exon 3 and 4. (B) Schematic of heterozygous SNP location on chromosome 18 in HCT116. Schematic was made with Phenogram created by Ritchie Lab (2012). (C) FISH on metaphase spreads stained with DAPI (blue) and hybridised to a chromosome 18 centromere probe (green) and 18q subtelomeric region probe (red). (D) Sanger sequencing traces of four heterozygous SNPs in HCT116. rs1056714 is located in the ZNF516 intron between crRNA-targeted exons 3 and 4. The rest of the SNPs are located distal to ZNF516 and towards the telomere. crRNA, CRISPR (clustered regularly interspaced short palindromic repeats) RNA; DAPI, 4′,6-diamidino-2-phenylindole; SNP, single-nucleotide polymorphism; UTR, untranslated region.