Overexpression of each of the 6 interfering TFs and 5 non-interfering TFs
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE29728 | GEO | 2011/06/07
SECONDARY ACCESSION(S): PRJNA153869
REPOSITORIES: GEO
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Project description:To clarify that the interfering effect in iPS induction was not because of extraordinary gene expression which was caused by overexpression of infected genes, transcriptional profile of the cells infected six genes were analysed using microarray analysis. As result, overexpression of each of the 6 interfering TFs in NSEB5-2C did not compromise transcriptional profile compared with the five non-interfering TFs. The cells were infected by retroviruses (carrying each ORF in pMXs-IRESNeo) at the titre in which around 95% of cells express the transgene. After 24hr of the infection, G418 was added to eliminate non-expressing cells. Each cells were harvested at 72hr after the infection.
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Project description:Overexpression of each of the 6 interfering TFs and 5 non-interfering TFs
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Project description:Although transcription factor(TF)s regulate differentiation-related processes, including dedifferentiation and direct conversion, functional interactions between TFs regulating these processes are not well understood. Here we show that TFs preventing dedifferentiation are able to induce direct conversion. Using a neural lineage cell line and a large number of TFs expressed in it, we found a subset of TFs whose overexpression strongly interfered with dedifferentiation triggered by a procedure to induce induced pluripotent stem cells (iPSC), through a maintenance mechanism of the cell-type-specific transcriptional profile. Strikingly, the maintenance activity of the interfering TF set was strong enough to induce the cell line-specific transcriptional profile when overexpressed in a heterologous cell type. In addition, TFs that interfered with dedifferentiation in hepatic lineage cells involved known TFs with induction activity for hepatic lineage cells. Our results suggest that dedifferentiation suppresses a cell-type-specific transcriptional profile, which is primarily maintained by a small subset of TFs capable of inducing direct conversion. We anticipate that this functional correlation might be applicable in various cell types, which may include cancer cells, and might facilitate identification of TFs with induction activity to understand differentiation and tumorigenesis Examination of binding of 3 transcription factors and histone modifications in Neural progenitor like cells.
2013-01-01 | E-GEOD-41252 | biostudies-arrayexpress
Project description:To clarify the gene expression profile of iHep, microarray analysis was performed using iHeps induced by 10 TFs (Foxg1, Lcor, Hnf3b, Hnf4a, Foxo6, Cdx2, Tcf1, Foxa3 ,Tcf2, Onecut1) and 9 TFs (Onecut1 was omitted from 10 TFs). Unsupervised hierarchical clustering indicated that iHep is expressing a global transcriptional profile more similar to that of HPCs rather than that of NPCs, and suggested that TFs present in the pool acted as inducing TFs. HPC (HB1 and HNG2) were established from fetal liver (E13.5) of C57BL6J and STOCK Tg(Nanog-GFP, Puro)1 Yam, respectively. iHeps were induced from NPC (NSBAg2, established from an ES cell line BAg73C2 carrying beta-geo knock-in allele in Afp) using retroviral vectors (pMXs without drug-selection markers) of 9 or 10 transcription factors. Three weeks after the infection, G418 was added and cultured for 1 week before the harvest. NSBAg2 and NSEB5-2C were used for the data of NPC. GSM396240 and GSM336010 were used for the data of ESC.
2011-06-07 | E-GEOD-29730 | biostudies-arrayexpress
Project description:To clarify the gene expression profile of iNPC, microarray analysis was performed using iNPCs induced by 6 TFs (Pax6, Hmga2, Etv6, Gatad2b, Nfxl1, and Esx1) and 5 TFs (Esx1 was omitted from 6 TFs). Unsupervised hierarchical clustering indicated that iNPC is expressing a global transcriptional profile more similar to that of NPCs rather than that of MEFs, and suggested that the TFs present in the pool acted as inducing TFs. iNPCs were induced from MEF using 6 or 5 transcription factors. iNPCs were induced from MEF (MEFSH, derived from mice carrying IRES-Hygro in Sox allele) using retroviral vectors (pMXs-IRESNeo) of 6 or 5 transcription factors. Four weeks after the infection, Hygromycin was added and cultured for 1 week before the harvest. NSBAg2 and NSEB5-2C were used for the data of NPC. GSM396240 and GSM336010 were used for the data of ESC.GSM651349 and GSM336011 were used for the data of MEF.
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Project description:Although transcription factor(TF)s regulate differentiation-related processes, including dedifferentiation and direct conversion, functional interactions between TFs regulating these processes are not well understood. Here we show that TFs preventing dedifferentiation are able to induce direct conversion. Using a neural lineage cell line and a large number of TFs expressed in it, we found a subset of TFs whose overexpression strongly interfered with dedifferentiation triggered by a procedure to induce induced pluripotent stem cells (iPSC), through a maintenance mechanism of the cell-type-specific transcriptional profile. Strikingly, the maintenance activity of the interfering TF set was strong enough to induce the cell line-specific transcriptional profile when overexpressed in a heterologous cell type. In addition, TFs that interfered with dedifferentiation in hepatic lineage cells involved known TFs with induction activity for hepatic lineage cells. Our results suggest that dedifferentiation suppresses a cell-type-specific transcriptional profile, which is primarily maintained by a small subset of TFs capable of inducing direct conversion. We anticipate that this functional correlation might be applicable in various cell types, which may include cancer cells, and might facilitate identification of TFs with induction activity to understand differentiation and tumorigenesis
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Project description:This SuperSeries is composed of the following subset Series: GSE29724: Unsupervised hierarchical clustering of iNPCs induced by 6 or 5 TFs GSE29726: The overexpression of Pax6 affects mesenchymal to epithelial transition (MET) pathway during iPS induction in NPC. GSE29728: Overexpression of each of the 6 interfering TFs and 5 non-interfering TFs GSE29729: Selection of NPC specific Transcription factors GSE29730: Unsupervised hierarchical clustering of iHPCs induced by 9 or 10 TFs Refer to individual Series
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Project description:To construct a candidate pool of NPC specific transcription factor (TF), we selected the genes expressed at relatively high levels in NPCs by comparing them to mixed RNA derived from multiple organs, eliminating housekeeping genes. As result, 160 putative NPC specific TFs were selected. We used three independent NPC lines (NSEB5-2C, NS195R, NS21) established from different ES cell lines (EB5, 195R, 21, respectively). Genes exhibiting relatively higher expression (1.5 fold) in NSEB5-2C and NS195R (or, NSEB5-2C and NS21) were selected by comparing with Mixture (consisting of equal microgram of RNAs from heart, kidney, thymus, spleen, liver, lung, d11 embryo). Gene Ontology (GO) terms ‘DNA binding’ and ‘transcription activity’ were used to select TFs.
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