PROJECT SUMMARY/ABSTRACT Genome-wide association studies (GWAS) and other genetic analyses consistently implicate non-coding cis- regulatory elements (CREs) in age-related macular degeneration (AMD). However, the contribution of CREs to the risk of developing AMD remains poorly understood because the function these elements has not yet been directly tested. The barrier for testing AMD-associated CREs is in part due to the challenge of developing experimentally validated model systems. CREs can be cell-type-specific and are not necessarily conserved between humans and model organisms. It therefore remains unclear which CREs may be studied in vivo in transgenic mice versus which require an in vitro human cell system. This gap in knowledge is a significant obstacle toward understanding the genetic regulation of normal human vision and to understanding the contribution of specific CREs to AMD. The long-term goal for our research is to understand how genetic variation within CREs shapes the structure and function of the retina and contributes to the risk of developing AMD and other disorders of vision. The focused objective of this proposal is to optimize and validate model systems for studying the function of AMD-associated CREs. The central hypothesis driving this work is that some AMD- associated CREs will be conserved between humans and mice due to their functional importance in vision. Other CREs may be human-specific due to their essential roles in the macula, a part of the retina that is not present in mice. In each of these cases, one or more optimal systems will be required to understand the function of each AMD-associated CRE. To test this hypothesis, we are pursuing the following specific aims: 1) Determine the functional conservation of AMD-associated CREs between humans and mice. 2) Test the function of a specific CRE that is known to be conserved between humans and mice from our preliminary studies in a newly developed mouse knockout model. 3) Test the requirement of human-specific CREs that are among the leading contributors to AMD risk, in induced human retinal tissue cultures. Together these experiments will enable discovery of genetic contributions to human vision and inherited visual diseases that have thus far been inaccessible using current methods.