In 2023, the National Academies held a series of facilitated workshops to gather information on progress to date on projects funded by the National Science Foundation’s (NSF’s) Understanding the Rules of Life (URoL) program. A fundamental goal of the URoL program is to identify the causal, predictive relationships that drive how life functions—a concept articulated as the “rules of life.” Principal investigators of the URoL program shared findings from their research and how those findings might contribute to identifying rules of life that are generalizable across fields and scales. Participants also discussed how they have incorporated multidisciplinary, systems-level approaches into their work. This booklet highlights the program’s focus areas and reach, major scientific breakthroughs, and lessons for research and education across scientific disciplines. This booklet is intended for policy and lay audiences interested in learning more about the goals and opportunities of NSF’s URoL program.