Functional analysis of human cardiac enhancers by tiling mutagenesis

To further determine the sequence features of active cardiac enhancers, 123 cardiac enhancers were functionally dissected by 10 bp tiling deletion-based mutagenesis MPRA in iPSC-CMs. Overall design: We performed systematic, tiling mutagenesis MPRA of 123 cardiac enhancers identified by 400 bp lentiMPRA. Each 400 bp region was represented as three fragments (F1, F2, F3), each containing 171 bp of genomic sequence. Each of these wild type sequences was tiled with 17 10 bp deletions.