TCF/LEF regulation of the topologically associated domain ADI promotes mESCs to exit the pluripotent ground state

Mouse embryonic stem cells (mESCs) can bemaintained in vitro in defined N2B27 medium supplemented with two chemical inhibitors for GSK3 and MEK (2i) and the cytokine leukemia inhibitory factor (LIF), which act synergistically to promote self-renewal and pluripotency. Here, we find that genetic deletion of the four genes encoding the TCF/LEF transcription factors confersm ESCs with the ability to self-renew in N2B27 medium alone. TCF/LEF quadruple knockout (qKO) mESCs display dysregulation of several genes, including Aire, Dnmt3l, and IcosL, located adjacent to each other within a topologically associated domain (TAD). Aire, Dnmt3l, and IcosL appear to be regulated by TCF/LEF in a beta-catenin independent manner. Moreover, downregulation of Aire and Dnmt3l in wild-type mESCs mimics the loss of TCF/LEF and increases mESC survival in the absence of 2iL. Hence, this study identifies TCF/LEF effectors that mediate exit from the pluripotent state.

Automated summary

The study demonstrates that genetic deletion of TCF/LEF transcription factors allows mESCs to self-renew in N2B27 medium independently of specific chemical inhibitors, possibly through the regulation of genes such as Aire, Dnmt3l, and IcosL. TCF/LEF are shown to regulate specific genes in a beta-catenin independent manner, influencing the pluripotent state of mESCs.