scRNAseq analysis of WT and tfeb-tfe3s triple knock-out zebrafish embryos at 4 days post fertilization

The transcription factors tfeb and tfe3 belong to the MiT/TFE gene family and are known as master regulators of lysosomal function and autophagy. In normal conditions, they are sequestered in the cytosol of the cells and their nuclear translocation is finely regulated in response to a huge array of cell signals; however, the role of these transcription factors during vertebrate development remains unclear. We used zebrafish to investigate tfeb and tfe3 role during embryo development and in adult organs. We generated knock-out (KO) alleles for all the zebrafish tfeb and tfe3s isoforms. Our data show that triple-KO (tfeb-tfe3a-tfe3b) animals are lethal, and these transcription factors are involved in pancreas and liver development. Wild-type (WT) and tfeb-tfe3a-tfe3b triple knock-out (TKO) embryos were collected at 4 days post fertilization (dpf), and GFP+ (neurons and pancreatic acinar cells) or dsRED+ (hepatocytes and pancreatic endocrine beta-cells) cells from dissociated embryos were isolated by Fluorescence-activated cell sorting (FACS) and analyzed by scRNA-seq.