Single-cell RNA-Seq Profiling of the Transcriptomic Response to RB Loss in the Retinoblastoma Cell of Origin

We applied single cell RNA-sequencing to define transcriptomic and cell cycle changes that follow RB loss in prenatal cone photoreceptor precursors, the cell of origin of retinoblastoma. We transduced intact retina and isolated cone precursors with a cone-specific H2B-cerulean fluorescent protein (CFP) reporter vector, and transduced prospectively isolated cones with YFP-marked lentivirus carrying either of two RB1 shRNAs or a SCR control shRNA. CFP+YFP+ cells were re-isolated by FACS for scRNA-seq at multiple time points over 15 days.