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. Overall design: 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.