BORIS is involved in transcriptional program of cancers. a The sequence recognized by ZFN and cleaved by Fok I is shown by black and red letters, respectively. b Surveyor assay (CEL-I) for ZFN-induced mutations in the BORIS gene. The proportions of wild-type and mutant alleles are shown as a table at the bottom of the gel. c Surveyor assay shows ten single-cell clones with the mutated BORIS locus. d Western blot shows the level of BORIS protein in wild type (wt) and ten mutant clones from panel (c). e K562 cells transfected with ZFN produced at least ten times less colonies in soft agar compared with control. f Western blot shows the level of BORIS protein in K562 before (0) and after phorbol 12-myristate 13-acetate (PMA) treatment. Wright–Giemsa staining analysis of K562 transfected with either control vector or ZFN in comparison with K562 treated with PMA for 3 days. Both transfection of K562 with ZFN and treatment with PMA showed morphologic changes related to megakaryocytic differentiation. h Upper panel: ChIP-seq and RNA-seq tracks show CTCF and BORIS occupancy at the WISP2 gene and dowregulation of WISP2 expression upon BORIS induction in MCF7 cells. Lower panel: RNA-seq RPKM values for the WISP2 gene expression. i Downregulation of inflammatory response pathway in response to stable BORIS expression in MCF7 cells. k ChIP-seq tracks show CTCF and BORIS occupancy at GAL3ST1, FOXA3, and PRAME loci in NHDF, OVCAR8, Delta47, and K562 cells. The tracks are labeled with the molecules against which antibodies were directed and cell lines used in ChIP-seq. RNA-Seq tracks demonstrate gene expression in BORIS-positive cells (K562) in contrast to BORIS-negative cells (NHEK and NHDF). l Quantitative PCR analysis of GAL3ST1, FOXA3, and PRAME gene expression in BORIS-positive cells (Testes, K562, OVCAR8, Delta47) and BORIS-negative cells (GM12878, NHDF, NHEK). (PPTX 1328 kb)