Allele-specific binding analysis of rs242561 by NRF2 ChIP tagmentation sequencing. Homo sapiens

We identified a SNP rs242561, located within a regulatory region of the MAPT gene (encoding microtubule-associated protein Tau). It was consistently occupied by NRF2/sMAF in multiple ChIP-seq experiments, and its strong-binding allele increased transactivation, and accorded higher mRNA levels in cell lines and human brain. To confirm the allele-specific binding, we conducted ChIP tagmentaion sequencing experiment in human lymphoblastoid cell line GM12763 which is heterozygous for rs242561. NRF2 ChIP DNA was isolated from GM12763 cells treated with Sulforaphane (SFN) in triplicates. The region containing rs242561 was amplified using primers, Fwd 5’-AGCCTTCCCTGTCCTTGATT-3’, Rev 5’-GGACCGAGCTTCCAGTCTAA-3’, and tagmentated using Tn5-based transposition for library construction. Libraries were sequenced on Illumina MiSeq. Overall design: NRF2 ChIP tagmentation sequencing of a 241bp intron region in MAPT gene using GM12763 treated with sulforaphane