Adolescent idiopathic scoliosis (AIS) affects about 3% of the population worldwide and is estimated to cost over 13 billion dollars in surgeries alone in the United States. The causes of AIS remain largely unknown. While mutations in genes leading to syndromic scoliosis (associated with other symptoms) have been discovered, the identification of mutations causing non-syndromic/isolated AIS (only AIS without any other symptoms), have been less successful. Several genome-wide association studies (GWAS) for scoliosis have identified AIS-associated SNPs in noncoding regions near genes that could be involved in AIS, suggesting that gene regulatory sequences, such as enhancers, could be a major cause of AIS. Using zebrafish and mouse enhancer assays, CRISPR/Cas 9 knockouts, RNA-seq and ChIP-seq we plan to characterize gene regulatory elements and pathways associated with AIS. Combined, our work will allow for the functional characterization of regulatory regions that can be a major cause of AIS and chart the regulatory landscape associated with this disease, thus providing additional candidate regions for mutation screening in individuals with AIS.