High-throughput mapping of regulatory DNA

摘要: Quantifying the effects of cis-regulatory DNA on gene expression is a major challenge. Here, we present multiplexed editing regulatory assay (MERA), high-throughput CRISPR-Cas9-based approach that analyzes functional impact genome in its native context. MERA tiles thousands mutations across ∼40 kb genomic space and uses knock-in green fluorescent protein (GFP) reporters to read out activity. Using this approach, obtain quantitative information contribution regions expression. We identify proximal distal elements necessary for four embryonic stem cell-specific genes. show consistent neighboring promoters unmarked (UREs) control but do not have typical enhancer epigenetic or chromatin features. compare nonfunctional genotypes at location base pair-resolution motifs elements.