The ability of single-cell RNA-Seq (scRNA-Seq) to measure the transcriptional basis of heterogeneity between individual cells is transforming biomedical research. It enables the precise ‘molecular dissection’ of the multiple cell types present within a complex tissue. This makes it possible to determine how a pathology affects each cell type, rather than just the overall effect upon the bulk tissue. As the technology matures and the range and scope of analytical tools expands, ever greater insights into cellular characteristics and their relationships to one another are becoming possible. However, each biological system presents unique challenges and customised experimental design and analyses are required. In this presentation I will outline recent and ongoing projects from my lab and collaborators which are applying scRNA-Seq to investigate the basis of the major eye diseases Diabetic Retinopathy, Age-related Macular Degeneration and Glaucoma. This work employs both in vivo models of the retina and in vitro cell culture systems with retinal endothelial, retinal pigment epithelial and lamina cribrosa cells. The results from a range of analytical approaches will be described, with examples chosen to help others maximise the pertinent information gained from their own experiments.

Learning Objectives:

1. Evaluate the potential of alternative technical and analytical single cell RNA-Seq approaches to provide relevant biological insights into a given model system.

2. Provide examples of how single cell RNA-Seq is being applied in vision science to understand the basis of eye diseases.