Determination of chemically-induced transcriptomic alterations in bladder oncogenesis using the developing zebrafish urinary system. This application is being submitted in response to the Notice of Special Interest (NOSI) identified as NOT-CA- 21-100. Manufacturing practices and chemical disposal have leached thousands of chemicals into our environment. Many of these chemicals, like bisphenol A (BPA), have concerning implications into multitudes of developmental anomaly and cancer incidences. Studying development expands our understanding of the cancer phenotype of mature tissues. Zebrafish are a well-established model in toxicogenomics - the study of interactions of chemicals and gene expression. They have recently been found to have a bladder which expands their potential for understanding bladder oncogenesis. We have designed a python-automated image analysis platform that sets the stage for a system that increases detection and analysis of chemical-induced phenotypes. The goal of this study is to determine the transcriptomic changes associated with developmental exposure to the BPA in an effort to better understand the oncogenic mechanisms associated with those exposures. Dr. Morhardt will carry out cloning experiments on a set of chemical-induced genes using CRISPR, inject those cloned genes into zebrafish larvae, and assess the changes associated with over- and under-expression of those genes. Zebrafish will also be exposed to BPA and examined for transcriptomic changes. Candidate genes from BPA exposure experiments will over- and underexpressed using CRISPR in a similar fashion to the initial CRISPR studies in aim 1. Genomic alternations will be compared with transcriptomic changes in TCGA database. These experiments will identify the transcriptomic mechanisms of chemically-induced developmental anomalies and create insights into chemically-induced oncogenesis