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MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons [RNA-Seq]

ABSTRACT: MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons [RNA-Seq]

PROVIDER: PRJNA962446 | ENA |

REPOSITORIES: ENA

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MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons

Project description:MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons

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Project description:Mutations in the methyl-DNA-binding protein MECP2 cause the neurodevelopmental disorder Rett syndrome (RTT). How MECP2 contributes to transcriptional regulation in normal and disease states is unresolved; it has been reported to be an activator and a repressor. We describe here the first integrated CUT&Tag, transcriptome, and proteome analyses using human neurons with wild-type and mutant MECP2 molecules. MECP2 occupies CpG-rich promoter-proximal regions in over four thousand genes in human neurons, including a plethora of autism risk genes, together with RNA polymerase II (Pol II). MECP2 directly interacts with Pol II, and genes occupied by both proteins showed reduced expression in neurons with MECP2 patient mutations. We conclude that MECP2 acts as a positive cofactor for Pol II gene expression at many neuronal genes that harbor CpG-islands in promoter-proximal regions, and that RTT is due, in part, to the loss of gene activity at these genes in neurons.

2024-04-26 | GSE230715 | GEO

MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons [RNA-Seq]

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MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons. [CUT&Tag]

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MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons [ATAC-seq]

Project description:MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons [ATAC-seq]

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MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons. [CUT&Tag]

Project description:MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons. [CUT&Tag]

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MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons

Project description:This dataset comprises 15 raw AP-MS files, each with an associated peak list, captured using a Vanquish Neo nanoLC system in tandem with an Orbitrap Eclipse mass spectrometer. To generate MECP2 hESC-reporter lines for wild type (WT) and various mutations of MECP2, including R133C, R168X, and R270X, we first used CRISPR/Cas9 to create MECP2 alleles carrying the green fluorescent protein (GFP) sequences in the endogenous gene. The R133C mutation was then introduced into the WT MECP2-GFP reporter line. Mutations R133C, R168X, and R270X are recognized as loss-of-function variants in MECP2 and are also identified as primary Rett syndrome-causing mutations. For efficient neuronal differentiation, a doxycycline (DOX)-responsive NGN2 construct was incorporated at their AAVS1 safe harbor locus. Upon the addition of DOX, homogenous populations of neurons were generated within three weeks from those four MECP2 hESC-reporter lines. Subsequently, GFP-pull down assay and AP-MS were performed using these WT MECP2-GFP neurons along with R133C-, R168X-, and R270X-mutant MECP2-GFP reporter neurons. Neurons expressing only the GFP tag served as a negative control. AP-MS analysis identified proteins interacting differently between WT and mutant MECP2 within human neurons.

2023-10-27 | MSV000093210 | MassIVE

MECP2 directly interacts with RNA polymerase II to modulate transcription in human neurons

Project description:This SuperSeries is composed of the SubSeries listed below.

2024-04-26 | GSE230716 | GEO