Cell-specific cis-regulatory elements and mechanisms of non-coding genetic disease in human retina and retinal organoids

Cis-regulatory elements (CREs) play a critical role in the development and disease-states of all human cell types. In the retina, CREs have been implicated in several inherited disorders. To better characterize human retinal CREs, we performed single-nucleus assay for transposase-accessible chromatin sequencing (snATAC-seq) and single-nucleus RNA sequencing (snRNA-seq) on the developing and adult human retina and on induced pluripotent stem cell (iPSC)-derived retinal organoids. These analyses identified developmentally dynamic, cell-class-specific CREs, enriched transcription-factor-binding motifs, and putative target genes. CREs in the retina and organoids are highly correlated at the single-cell level, and this supports the use of organoids as a model for studying disease-associated CREs. As a proof of concept, we disrupted a disease-associated CRE at 5q14.3, confirming its principal target gene as the miR-9-2 primary transcript and demonstrating its role in neurogenesis and gene regulation in mature glia. This study provides a resource for characterizing human retinal CREs and showcases organoids as a model to study the function of CREs that influence development and disease.

Automated summary

Cis-regulatory elements (CREs) are crucial for the development and disease-states of human cell types, including the retina. This study used single-nucleus assay and sequencing techniques to characterize human retinal CREs in developing and adult retina as well as retinal organoids. The results provide insights into the dynamic nature of cell-specific CREs and their potential target genes, and confirm the function of a disease-associated CRE in neurogenesis and gene regulation.