Investigating the role of human UTF1 in reprogramming, self-renewal and differentiation using a CRISPR/Cas toolbox

Abstract

Stem cells are self-renewing cells present at the apex of the lineage hierarchy and, therefore, serve as the founder cells during organismal development. Embryonic Stem Cells (ESCs) are pluripotent cells that can differentiate into all the cell types belonging to the three germ layers: ectoderm, mesoderm, and endoderm. However, human ESCs are not considered ideal for cell therapy applications because of ethical issues and their inability to be used for autologous therapy. Circumventing these limitations, a ground- breaking study was published in 2006, in which pluripotency was induced in terminally differentiated cells (fibroblasts) by the introduction of a cocktail of transcription factors, namely OCT3/4, SOX2, KLF4, and c-MYC (popularly called Yamanaka factors) in mouse fibroblasts to generate induced Pluripotent Stem Cells (iPSCs). Subsequently, the first human iPSCs were reported from fibroblasts using retroviral and lentiviral transduction of reprogramming factors. Since its inception, various reprogramming approaches and combinations of reprogramming factors have been explored to generate iPSCs with higher efficiency and quality. Among these, a pluripotent cell-specific transcription factor Undifferentiated embryonic cell Transcription Factor 1 (UTF1) was believed to be a promising factor for the generation of quantitatively and qualitatively better human iPSCs due to its high expression in pluripotent stem cells. Therefore, our aim was to elucidate the role of human UTF1 in the generation and maintenance of human iPSCs.