A Comparative Analysis of Single-Cell Transcriptome Identifies Reprogramming Driver Factors for Efficiency Improvement

摘要

Terminally differentiated somatic cells can be reprogrammed into a totipotent state through somatic cell nuclear transfer (SCNT). The incomplete reprogramming is the major reason for developmental arrest of SCNT embryos at early stages. In our studies, we found that pathways for autophagy, endocytosis, and apoptosis were incompletely activated in nuclear transfer (NT) 2-cell arrest embryos, whereas extensively inhibited pathways for stem cell pluripotency maintenance, DNA repair, cell cycle, and autophagy may result in NT 4-cell embryos arrest. As for NT normal embryos, a significant shift in expression of developmental transcription factors (TFs) , , , and was observed. Compared with pluripotent gene being activated only in NT 2-cell, , , and had major expression waves in normal development of both NT 2-cell and 4-cell embryos. Additionally, / and had been confirmed as key markers in NT 2-cell and 4-cell embryos, respectively. Histone acetylases , , and were co-activated in NT 2-cell and 4-cell embryos to facilitate normal development. as a key driver functions with and to improve the efficiency of NT reprogramming. Taken together, our findings provided an important theoretical basis for elucidating the potential molecular mechanisms and identified reprogramming driver factor to improve the efficiency of SCNT reprogramming.