UCSF

Most of the human genome does not encode proteins but instead contains a vast array of non-coding sequences that play crucial roles in gene regulation. Understanding these regulatory sequences, especially in neuronal contexts, is essential for uncovering brain function and development complexities. In this work, I utilized high-throughput assays alongside the mouse as a model to examine neuronal non-coding regulatory sequences. In Chapter 1, I tested thousands of candidate regulatory elements using Massively Parallel Reporter Assays (MPRA). We further validated strong candidates using mouse transgenic assays to assess the enhancer activity in vivo. Our combined approach of MPRA and mouse transgenic assays revealed complementary information on enhancer activity, highlighting the strengths and limitations of each method. In Chapter 2, I focused on functionally characterizing the regulatory network of the oxytocin receptor. The oxytocin receptor is a key regulator of social behavior. We identified seven candidate regulatory elements using comparative and functional genomics tools. We further validated the enhancer activity of the strongest candidate regulatory element using stable mouse transgenic lines. We determined the candidate regulatory element to have enhancer activity in the mouse olfactory bulb at post-developmental stages. This comprehensive study underscores the intricate regulatory landscapes that govern neuronal functions and showcases the power of integrating high-throughput screening with in vivo validation to unravel these complexities.