Reprogramming the CNS of the Proto-Vertebrate Ciona intestinalis Using Single-Cell Transcriptomics

We applied single cell RNA seq with 10x genomics platform to study co-expression of both Ptf1a and Meis leading to the wholesale transformation of the entire CNS into Coronet cells. Our study identify a cocktail of regulatory determinants for the efficient specification of Coronet cells. We injected Ptf1a>β2tubulin construct into Ciona oocytes, fertilized and let them developed into middle tailbud stage along with control. We also injected Ptf1a and Meis>β2tubulin construct into Ciona oocytes, fertilized and let them developed into late tailbud stage along with control. Embryos were dissociated into single cells and loaded into 10x platform for single cell RNA-seq. Reads are aligned to Ciona genome with Cell Ranger pipeline.