ZBTB10 binds the telomeric variant repeat TTGGGG and interacts with TRF2 [DNA-Seq validation of KO]. ZBTB10 binds the telomeric variant repeat TTGGGG and interacts with TRF2 [DNA-Seq validation of KO]

In order to investigate changes upon ZBTB10 depletion, we used CRISPR/Cas9 to induce a ZBTB10 knockout in HEK293 and U2OS cells. Overall design: ZBTB10 knockouts were generated using three different guide RNAs targeting the second exon of the ZBTB10 gene. HEK293 and U2OS cells were selected in 750 ng/mL puromycin after transfection and gDNA was extracted. A circa 1100 bp long target sequence around the cut locus was amplified by PCR and Cas9 activity was tested in a T7 endonuclease assay. After confirming Cas9 activity, single cell sorting was performed and cells were expanded to screen for ZBTB10 knockout clones by Western blot using an anti-ZBTB10 antibody (Supplementary Table 1).