Genome-wide analysis of iPSC-derived neurons with EGFP or TERT transduction
Ontology highlight
ABSTRACT: Analysis of TERT-dependent global gene expression changes in iPSC-derived human AD neurons. The hypothesis tested in the present study was that TERT influences the regulation of gene expressions in the iPSC-derived human neurons. Results provide important information of the response of iPSC-derived neurons in presence and absence of TERT induction.
Project description:Analysis of TERT-regulated global gene expression changes. The hypothesis tested in the present study was that neuronal TERT influences the regulation of gene expressions. Results provide important information of the response of in vivo mouse neurons to physiological TERT induction.
Project description:sample 666823 and 666824 control IgG pulldown in human iPSC derived APPDp neurons
sample 666825 and 666826 TERT pulldown in human iPSC derived APPDp neurons
Project description:Compared the global gene expression profiles of HD- and CON-iPSC-derived neurons We used microarrays to detail the global programme of gene expression for comparing the global gene expression profiles of HD- and CON-iPSC-derived neurons and facilitating studies of medium spiny neurons (MSN)-degenerative processes of Huntington's Disease (HD). By using a step-wise in vitro differentiation protocol combining EB formation, neural induction by small molecules, treatment with inhibitors of the TGFß pathway (SB431542) and the BMP pathway (LDN193189), and mechanical isolation/purification of neural progenitors and neurons, we induced 60-70% of control iPSCs or HD-iPSCs to differentiate into GABA- and DARPP-32- double positive neurons.
Project description:The goal of this study is to understand how the specific TERT inhibitor BIBR1532 preconditioning regulates transcriptional reprogramming in mouse neuronal cells. We use CUT-Tag approach to comprehensively map the genomic binding sites of TERT in primary neurons and determine the alteration of is binding profile after BIBR1532 preconditioning. By obtaining over 10 million reads per sample, we generated genome-wide TERT chromatin-binding maps of mouse primary neurons. Under normal conditions, we show that more than 50% of the TERT binding sites were located at the promoter regions, many of them are previously unknown genomic loci. Intriguingly, BIBR1532 preconditioning significantly alters TERT-chromatin binding profile. Out of the total promoter binding sites by TERT, 30.4% (2,837 peaks) and 31.7% (2,954 peaks) are respectively upregulated and downregulated in TERT-binding after BIBR1532 preconditioning. Interestingly, we find that BIBR1532-preconditioned neurons show significant upregulation of promoter binding of TERT to the mitochondrial antioxidant genes. Together, these results identify the previously unknown TERT-binding sites in the mouse primary neurons and demonstrate that BIBR1532 preconditioning confers neuronal ischemic tolerance through TERT-mediated transcriptional reprogramming.
Project description:Compared the global gene expression profiles of HD- and CON-iPSC-derived neurons We used microarrays to detail the global programme of gene expression for comparing the global gene expression profiles of HD- and CON-iPSC-derived neurons and facilitating studies of medium spiny neurons (MSN)-degenerative processes of Huntington's Disease (HD).
Project description:TERT is an essential protein component of telomerase, a ribonuclearprotein complex that protects chromosomal ends. Ectopic expression of TERT in mouse skin activates hair follicle stem cells and induces active growth phase of hair cycles, called anagen. This activity of TERT is independent of its reverse transcriptase function, indicating that this is a non-telomeric function of TERT. We performed time-course microarray analysis using conditional bi-transgenic mice (iK5-TERT) to understand the mechanism of this novel non-telomeric function of TERT. Experiment Overall Design: Hair follicles of iK5-TERT mice at postnatal day 60 is in TERT-induced anagen. To those mice, we injected either PBS, to maintain TERT transgene expression, or doxycycline, to rapidly extinguish transgene expression. We then took time-course dorsal skin biopsies (0,6,12,24 hr) of these mice and analyzed differential gene expressions using Affymetrix Genechips and various bioinformatic tools, including SAM, to discover TERT-regulated genes.