library_construction_protocol: After dissociation cells were counted by FACS (no sorting). Cell concentration was determined using a TC20 automated cell counter (BioRad) Cells grown in 2iLIF on gelatin coated plates and transferred to SerumLIF medium on gelatin for 86h Mouse embryonic stem cells, transfected with gRNAs CACACTCAAGGATCCACAGG and GCCACAGATGGTGTCCGCGG,  Dax1 mutant clone Cells for the samples were loaded into channel of a Chromium Single Cell A Chip (10X genomics) channels. Reverse-transcription, cDNA pre-amplification (13 cycles) and library preparation were performed according to the manufacturer instructions (Chromium Single Cell 3â€² Library & Gel Bead Kit v3, 10X genomics).