Analysis of CHD non-coding DNVs for their impact on cardiac cis-regulatory element activity

6761 CHD non-coding DNVs were prioritized from 749 unexplained CHD trios and functionally assessed in iPSC-CMs using lentiMPRA. We synthesized 6761 reference (REF) and variant (ALT) pairs, comprising 171 bp of genomic sequence centered on the DNVs. The CHD MPRA library also included 865 negative controls (ATAC-seq peaks in iPSCs and not iPSC-CMs) and 613 additional oligos derived from the 18 enhancers with the most subregion activity in the mutagenesis MPRA. The 15000 pooled oligos were synthesized following the same barcoded design as used for tiling mutagenesis.