Effect of Dnmt3a/3b transient silencing in WT iPSCs and comparison with miR-203 tKI iPSCs

The balance between pluripotency and differentiation is critical during development and regeneration. miR-203 is a microRNA previously involved in differentiation of different tissues as well as in tumor suppression in multiple malignancies. We have shown that miR-203 is able to promote differentiation of embryonic stem (ES) and induced pluripotent stem (iPS) cells without decreasing pluripotency. We have observed that transient expression of miR-203 significantly improves the efficiency of ES/iPS cells in the generation of quimeras and tetraploid complementation assays, in addition to inducing complex embryo-like structures when these pluripotent cells are injected in mice. Mechanistically, we have shown that miR-203 mediates such effects, at least in part, by modulating the levels of de novo DNA methyltransferases. In the present RNAseq we have transiently silenced the levels of DNMT3a/3b in order to compare their transcriptomic profile with that observed in PSCs transiently exponed to miR-203. Overall design: The general idea was to analyze the transcriptomic profile of WT iPSCs transiently transfected with siRNA control, Dnmt3a,Dnmt3b or both and compare their transcriptomic signatures with those observed in mir-203 tKI iPSCs. As described previously, miR-203 tKI IPSCs were transiently induced by doxycycline treatment for 5 days, and samples were collected 2 weeks after dox withdrawal.