Fig. 1: Structure of AATK and hypermethylation of its CpG island promoter in cancer.

A AATK is characterized by a tyrosine kinase domain (NCBI tool for conserved domain search) [76]. B AATK is located on chromosome 17q25.3. A 537 bp CpG island (CGI) (green) overlaps the transcription start site [40]. The frequency of the individual CpG sites is depicted (black lines). For the methylation analyses via combined bisulfite restriction analysis (CoBRA) and pyrosequencing, a 237 bp PCR product is generated (gray bar). Two TaqI restriction sites for CoBRA (red), three CpGs for pyrosequencing (purple), and two CpGs (cg26717786 and cg15665342) for Illimuna450k array (blue) are marked. For epigenetic editing, four gRNAs (#1–#4) covering the CGI were generated (light green). C A panel of 14 cancer cell lines with three head and neck, two breast, two thyroid, one sarcoma, one ovary, two brain cancer cell lines, and an in vitro methylated (ivm) sample were analyzed via CoBRA. PCR products were mock (−) and TaqI (+) digested.