Human donor eyes, consented for research, were obtained from the Eye hospital and were approved by the Ethics Committee of Optometry Hospital affiliated to Wenzhou Medical University (2022-121-K-92-01). iPSCs were passaged onto Matrigel-coated dish in retinal differentiation medium (RDM, DMEM/F12 with 1% B27 (Gibco, 12587010), 1% N2 (Gibco, 17502048) and 1×NEAA (Gibco, 11140050). 10 mM nicotinamide (Selleck, S1899, Houston, Tx, USA), 10 ng/ml IGF1 (Sino Biological, 10598-HNAE, Beijing, China), 10 ng/ml bFGF (Sino Biological, 10014-HNAE), 1 M A-83-01 (Selleck, S7692) and 1 M IWR-1 (Selleck, S7086) were added to RDM for 0-4 days. 10 mM nicotinamide and 10 ng/ml bFGF were replaced by 100 ng/ml Activin A (Sino Biological, 10429-HNAH) and the concentration of IGF1 increased to 20 ng/ml, and A-83-01 was not added for 4-6 days. IGF1 and IWR-1 were replaced by 1 M SU5402 (Selleck, S7667) for 6-8 days. 1 M CHIR99021 (Selleck, S1263) was added to RDM for 8-14 days. From days 14, cells were maintained in RPE supporting medium (RSM, DMEM/F12 with 15% knockout serum replacement, 5% FBS, 1×NEAA and 10 mM nicotinamide) until they matured and formed pigmented monolayers. iPSC-RPE cells were passaged with EDTA (Gibco, 25200072) and were replaced on the Matrigel-coated dish and at P3 were used in this study. Histone modifications of ARPE-19, iPSC-RPE, hRPE and iPSC were captured by CUT&Tag using hyperactive universal CUT&Tag assay kit for Illumina (Vazyme, TD903, Nanjing, China). 50,000 viable cells were collected for each library and incubated with concanavalin A-coated magnetic beads. The beads binding cells were captured by H3K4me1 (CST, 5326, Danvers, MA, USA), H3K4me3 (CST, 9751), H3K27ac (CST, 8173), H3K27me3 (CST, 9733) antibodies, then incubated with hyperactive pA/G-Transposon. The tagmented DNA was purified and PCR amplificated for sequencing.