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Next Generation Sequencing of wild-type HEK293 cells compared to STAT5b knock-out HEK293 cells using Crispr/Cas9

ABSTRACT: Next Generation Sequencing of wild-type HEK293 cells compared to STAT5b knock-out HEK293 cells using Crispr/Cas9

PROVIDER: PRJNA732661 | ENA |

REPOSITORIES: ENA

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Next Generation Sequencing of wild-type HEK293 cells compared to STAT5b knock-out HEK293 cells using Crispr/Cas9

Project description:During kidney diseases, diverse tissue-specific pathways can regulate kidney injury and prognosis. In our study, we focused on STAT5, a member of the Janus Kinase/ Signal Transducer and Activator of Transcription (JAK/STAT) pathway, classically described in immune cells which has been recently described in intrinsic kidney cells. STAT5 activation is detected in both human podocytes and tubular cells in FSGS and ATI, respectively. Additionally, STAT5 deficiency in either glomerular or tubular epithelium aggravates the functional and structural alterations in a range of experimental models of glomerular or tubular disease. Using RNA sequencing, we show here that STAT5b inactivation in HEK-293 cells resulted in dysregulation of multiple metabolic pathways, including glycolysis and oxidative phosphorylation. In conclusion, activating renal epithelial STAT5 represents a new therapeutic avenue with the potential for a range of beneficial effects in kidney disease.

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Landscapes of TET1 and STAT5B binding on DNA across the entire genome in PDX2 B-ALL cells

Project description:Here we report both TET1-WT and catalytically dead mutant TET1 (TET1-MUT) directly interact with STAT5B in PDX2 B-ALL cells. To study the landscape of TET1 and STAT5B binding in whole genomic DNA and test if the STAT5B binding signal is affected by TET1, we conduct the ChIP-seq assay with flag antibody (TET1-WT, TET1-MUT) and STAT5B antibody (sgNS or sgTET1) respectively. The results showed that TET1 bound peaks overlap with STAT5B very well and the tag density of STAT5B on target genes decreases in TET1-impeded cells. We conclude that TET1 recruit STAT5B to the promoters of its target genes and further promote the target genes’ transcriptions.

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Sex-dependent and STAT5b-dependent Liver Gene Expression

Project description:A series of dual-channel gene expression profiles obtained using Rosetta/Merck Mouse TOE 75k microarrays was used to examine the sex-dependent and STAT5b-dependent differences in gene expression in adult mouse liver. This series is comprised of 4 pools of 3 randomly chosen independent wildtype male and female mouse liver cDNA samples and 4 pools of 3 randomly chosen independent STAT5b-deficient male and female mouse liver cDNA samples, totaling 16 pools. The pools were paired randomly to generate 4 comparisons of M-WT:F-WT, M-WT:M-KO, F-KO:F-WT, and F-KO:M-KO. Comparison of the set of sex-dependent genes with the set of genes responsive to the loss of STAT5b in males shows that 75% of the sex-specific genes were also regulated by STAT5b in males. Only 20% of the sex-specific genes retained sex-specificity in the absence of STAT5b, indicating a large role for STAT5b in sex-specific liver gene expression. Keywords: genetic knockout and sex response

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