Functional dissection of human cardiac enhancers with hiPSC-CMs and lentiMPRA

To assess human cardiac enhancer activity, 400 bp candidate cardiac enhancers and controls were tested in iPSC-CMs using lentiMPRA. Overall design: We applied lentiMPRA to measure the enhancer activity of 2891 candidate human cardiac enhancers and 859 negative controls. iPSC-CMs were transfected with the lentiMPRA library at day 10 or 17 of iPSC-CM differentiation, and cells were collected 7 days later. A region of the 3'UTR of the reporter gene containing the candidate CREs was amplified from RNA and genomic DNA and sequenced.