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Comparison of genomic binding profile for RNA polyII in control and Rex1 RNAi hESCs (H9 cell line)

ABSTRACT: To understand if rex1 is directly involved in CFIm25-regulated transcription at the transcription level, we carried out ChIP-seq analysis for RNAPII in control and rex1 RNAi cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE178193 | GEO | 2022/04/21

REPOSITORIES: GEO

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Comparison of genomic binding profile for RNA polyII in control and CFIm25 KD hESCs (H9 cell line)

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Comparison of genomic binding profile for CFIm25 in control and CFIm25 KD hESCs (H9 cell line)

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Comparison of genomic binding profile for RNA polyII in control and Rex1 RNAi hESCs (H9 cell line)

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Comparison of genomic binding profile for CFIm25 in control and CFIm25 KD hESCs (H9 cell line)

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poly(A+)-seq profiling of gene expression in control H9 and (CFIm25 KD+ 3Flag-CFIm25 overexpression) H9 cell lines

Project description:We carried out CFIm25 KD experiments in H9 cell line (hESCs). After poly(A+)-seq, we found the expression of hundreds of genes were changed. To investigate if the effect is a direct or indirect effect of CFIm25 KD, we next carried out 3Flag-CFIm25 overexpression in CFIm25 KD cells for gene-rescue experiments. After we obtained this cell line, here we performed poly(A+)-seq in control H9 and (CFIm25 KD+ 3Flag-CFIm25 overexpression)H9 cell lines. The goal of this experiment is to investigate if CFIm25 overexpression could rescue the gene expression phenotype caused by CFIm25 KD.

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Comparison of genomic binding profile for RNA polyII in control and CFIm25 KD hESCs (H9 cell line)

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Ser7 of RNAPII-CTD facilitates heterochromatin formation by linking ncRNA to RNAi

Project description:Some long noncoding RNAs (ncRNAs) transcribed by RNA polymerase II (RNAPII) are retained on chromatin, where they regulate RNAi and chromatin structure. The molecular basis of this retention remains unknown. We show that in fission yeast serine 7 (Ser7) of the C-terminal domain (CTD) of RNAPII is required for efficient siRNA generation for RNAi-dependent heterochromatin formation. Surprisingly, Ser7 facilitates chromatin retention of nascent heterochromatic RNAs (hRNAs). Chromatin retention of hRNAs and siRNA generation requires both Ser7 and an RNA-binding activity of the chromodomain of Chp1, a subunit of the RNA-induced transcriptional silencing (RITS) complex. Furthermore, RITS associates with RNAPII in a Ser7- dependent manner. We propose that Ser7 promotes cotranscriptional chromatin retention of hRNA by recruiting the RNA-chromatin connector protein Chp1, which facilitates RNAi-dependent heterochromatin formation. Our findings reveal a function of the CTD code: linking ncRNA transcription to RNAi for heterochromatin formation.

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