Generation of a human iPSC line with EGFP knock-in at the ATRX locus for visualization of chromatin remodeler dynamics and nuclear condensate formation

ATRX is a chromatin-remodelling protein associated with neurodevelopmental disorders and gliomas, forming distinct nuclear foci associated with chromatin organization and phase-separated nuclear compartments. To visualize ATRX dynamics in live cells, we generated a human iPS cell (hiPSC) line with EGFP knocked in at the ATRX N-terminus using CRISPR/Cas12a-mediated genome editing. The modified hiPSCs retained pluripotency, normal karyotype, and trilineage differentiation potential. EGFP-ATRX localized to discrete nuclear foci, consistent with the phase-separated condensates described for ATRX, enabling real-time visualization of its chromatin association. This reporter line offers a valuable tool for studying ATRX function during differentiation and chromatin architecture development.