Project description:Reprogramming to induced pluripotency induces the switch of somatic cell identity to induced pluripotent stem cells (iPSCs). However, the mediators and mechanisms of reprogramming remain largely unclear. To elucidate the mediators and mechanisms of reprogramming, we used a siRNA mediated knockdown approach for selected candidate genes during the conversion of somatic cells into iPSCs. We identified Tox4 as a novel factor that modulates cell fate, using reprogramming efficiency towards iPSCs as an assay. We found that Tox4 is needed early in reprogramming to efficiently generate early reprogramming intermediates, irrespective of reprogramming conditions used. Tox4 enables proper exogenous reprogramming factor expression and the closing and opening of putative somatic and pluripotency enhancers early during reprogramming, respectively. We show that TOX4 protein assembles into a high molecular form. Moreover, Tox4 is also required for the efficient conversion of fibroblasts towards the neuronal fate, suggesting a broader role of Tox4 in modulating cell fate. Our study reveals Tox4 as a novel transcriptional modulator of cell fate that mediates reprogramming from the somatic state to the pluripotent and neuronal fate.
Project description:Reprogramming to induced pluripotency induces the switch of somatic cell identity to induced pluripotent stem cells (iPSCs). However, the mediators and mechanisms of reprogramming remain largely unclear. To elucidate the mediators and mechanisms of reprogramming, we used a siRNA mediated knockdown approach for selected candidate genes during the conversion of somatic cells into iPSCs. We identified Tox4 as a novel factor that modulates cell fate, using reprogramming efficiency towards iPSCs as an assay. We found that Tox4 is needed early in reprogramming to efficiently generate early reprogramming intermediates, irrespective of reprogramming conditions used. Tox4 enables proper exogenous reprogramming factor expression and the closing and opening of putative somatic and pluripotency enhancers early during reprogramming, respectively. We show that TOX4 protein assembles into a high molecular form. Moreover, Tox4 is also required for the efficient conversion of fibroblasts towards the neuronal fate, suggesting a broader role of Tox4 in modulating cell fate. Our study reveals Tox4 as a novel transcriptional modulator of cell fate that mediates reprogramming from the somatic state to the pluripotent and neuronal fate.
Project description:This study determined whether human adipose stem cells (hASCs) could be reprogrammed into induced pluripotent stem (iPS) cells using the 4 factors OCT4, SOX2, KLF4, and cMYC. We showed successfull reprogramming could be achieved in which fat/adipose cells were able to assume embryonic-like phenotypes after lentiviral transduction with the 4 factors. Further, we believe these fat cells are intrinsically better-suited for reprogramming compared to fibroblasts, and result in higher yields of iPS cells. Microarray study was performed in duplicates (hASCs, iPS cells derived from hASCs, and hESCs). Reference control was pooled RNA taken from H9 hESCs, embryoid bodies, and some differentiated cell types such as endothelial cells.
Project description:This SuperSeries is composed of the following subset Series: GSE26428: Effect of Glis1 on human iPS cell generation GSE26429: Promotion of Direct Reprogramming by Glis1 GSE26430: Effect of Glis1, Dmrtb1, and Pitx2 on mouse iPS cell generation Refer to individual Series
Project description:This study determined whether human adipose stem cells (hASCs) could be reprogrammed into induced pluripotent stem (iPS) cells using the 4 factors OCT4, SOX2, KLF4, and cMYC. We showed successfull reprogramming could be achieved in which fat/adipose cells were able to assume embryonic-like phenotypes after lentiviral transduction with the 4 factors. Further, we believe these fat cells are intrinsically better-suited for reprogramming compared to fibroblasts, and result in higher yields of iPS cells.
Project description:We reprogrammed human CD34+ cells from cord blood using a lentiviral vector encoding OCT4, SOX2 and KLF4.We collected RNA from parental CD34+ cells (3samples), reprogramming timepoints (9 timepoints), iPS clones derived from this experiment (6 clones), and human ES cell lines (9 samples). All samples were sequenced at 100bp reads. Endogenous retroelement expression during reprogramming