The regulation of gene expression is essential for ensuring normal developmental processes. Dysregulation of gene expression programs can have profound effects on cellular functions and underlies many diseases. Since the monumental discovery of RNAPII just over 50 years ago, it has become increasingly clear that gene control is a dynamic and highly regulated process. This meeting will explore unexpected layers in the regulation of gene expression. We will discuss new advances in the roles of regulatory factors, cis elements, and epigenetic mechanisms. New insights into genome transcription and regulation will be revealed through structural biology, with significant advances in Cryo-EM, and functional genomics in eukaryotic systems. This meeting will also include approaches for advancing our understanding of the evolution of gene regulation. Finally, this meeting will bring together innovative approaches and technologies including single cell measurements of gene expression, cutting-edge imaging, and long read sequencing to uncover secrets of the regulatory genome.