Endothelin-1 (EDN1) plays a critical role in regulation of blood pressure through its biological activity as a potent vasoconstrictor. The Gumz lab discovered that cultured endothelial cells express an RNA transcript that extends in an antisense orientation relative to the direction of EDN1 transcription, and we hypothesize that this “antisense transcript” functions to regulate expression of EDN1 as a long, non-coding RNA (lncRNA). Inspection of the ENCODE data base showed a ~350 bp DNaseI hypersensitive site directly 3’ to EDN1 containing numerous genetic and epigenetic signatures consistent with function as a promoter. My research focuses on using CRISPR/Cas9 gene editing technology to delete this supposed promoter sequence in cultured cells and subsequently determine the effects this manipulation has on antisense transcript and EDN1 gene expression. I am currently designing sgRNAs to target each flank of the EDN1 antisense promoter and I will screen cells for bi-allelic deletion by PCR of genomic DNA from selected cell clones. Following successful completion of gene editing, I will assess antisense and EDN1 transcript expression by quantitative PCR using strand-specific primers.