Project description:Partial exon deletion in Trpm7 gene was confirmed by lack of transcripts according to the RNA sequencing in two stem cell clones. For this mice were genetically modified by partial deletion of exon 2 and embryonic stem (ES) cell lines were generated. Isolated RNA of two ES clones was sequenced with Illumina NextSeq 500 Sequencer, which generated 75 bases long ‘single-end’ reads.
Project description:TRPM7 is a ubiquitous ion channel and kinase, a unique ‘chanzyme’, required for proper early embryonic development. In order to assess the effects of TRPM7 activity on cellular gene expression, mouse embryonic stem cells with TRPM7 gene deletion (TRPM7-/- mESC) were established. By using microarray analysis, we identified genes with transcription significantly different in TRPM7-deficient mESC.
Project description:TRPM7 is a ubiquitous ion channel and kinase, a unique M-bM-^@M-^XchanzymeM-bM-^@M-^Y, required for proper early embryonic development. In order to assess the effects of TRPM7 activity on cellular gene expression, mouse embryonic stem cells with TRPM7 gene deletion (TRPM7-/- mESC) were established. By using microarray analysis, we identified genes with transcription significantly different in TRPM7-deficient mESC. Total RNA was extracted from cells using Qiagen RNAeasy Plus mini Kit. Triplicate samples were made for each wild type clone 3 (WT3) and and TRPM7-/- clone 9 (KO9) mESC
Project description:Muscle satellite cells (MuSCs), skeletal muscle-resident stem cells, are crucial for regeneration of myofibers. Molecular factors such as cations that trigger the transition of MuSCs from a quiescent to an active state remain largely unidentified. In this study, we identified TRPM7, a Mg2+-permeable ion channel, as a critical determinant for myofiber regeneration. We investigated gene profiles of Trpm7-deficient MuSCs to understand the role of TRPM7 during myogenesis. Our results suggest that TRPM7 governs the myogenesis, cell-cycle progression and mitochondrial biosynthesis in satellite cells to regulate crucial events (ie, activation, proliferation, and myogenesis) during skeletal muscle regeneration.
Project description:Analysis of the effect of shRNA-mediated knockdown of TRPM7 on gene expression levels in SH-SY5Y and SH-EP2 human neuroblastoma cells. Results were used for the identification of neural-crest-associated transcription factors that were affected by TRPM7 knockdown. Total RNA isolated from SH-EP2 and SH-SY5Y human neuroblastoma cells transduced with a scrambled shRNA (control) or TRPM7 shRNA, experiment performed in duplicate.
Project description:The functional activity of TRPM7 is essential for cell viability and growth, and its expression is up-regulated in certain pathological conditions. In order to assess the effects of TRPM7 activity on cellular gene expression, inducible HEK293 cell-lines harboring the wild-type mouse TRPM7 and a mutant lacking the kinase domain were established. By using microarray analysis, we identified genetic profiles altered in transcription significantly and specifically by the expression of the functional TRPM7 channel. Overexpression of TRPM7 channel in HEK cells induce the cell damage and cell death. So we used tetracycline-inducible system for channel expression. However, since the treatment of inducer itself could also affect the transcription profile, we established another cell-line harboring a nonfunctional TRPM7 mutant (TRPM7DKD) as a control in which the entire kinase domain at the C-terminus was deleted. The wild-type and the non-functional TRPM7 channels were induced transiently, and the comparative changes in cellular transcription were investigated