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Genome binding/occupancy profiling of ALKBH5 by high throughput sequencing [Cut&Tag]

ABSTRACT: Genome binding/occupancy profiling of ALKBH5 by high throughput sequencing [Cut&Tag]

PROVIDER: PRJNA1044792 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26942650_1.fastq.gz	Fastqsanger.gz
	SRR26942650_2.fastq.gz	Fastqsanger.gz
	SRR26942651_1.fastq.gz	Fastqsanger.gz
	SRR26942651_2.fastq.gz	Fastqsanger.gz

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Genome binding/occupancy profiling of ALKBH5 by high throughput sequencing [Cut&Tag]

Project description:Alkylation repair homolog protein 5 (ALKBH5) binds to RNA and mediates RNA m6A demethylation. However, whether ALKBH5 binds to DNA is largely unknown. In this study, our CUT&Tag assay identified 12333 peaks in ALKBH5-overexpressing Hepa1-6 cells, indicating that ALKBH5 can bind to DNA.

2025-02-27 | GSE248568 | GEO

Genome binding/occupancy profiling of ALKBH5 by high throughput sequencing [CUT&RUN]

Project description:Genome binding/occupancy profiling of ALKBH5 by high throughput sequencing [CUT&RUN]

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Genome binding/occupancy profiling of ALKBH5 by high throughput sequencing [CUT&RUN]

Project description:Alkylation repair homolog protein 5 (ALKBH5) binds to RNA and mediates RNA m6A demethylation. However, whether ALKBH5 binds to DNA is largely unknown. In this study, our CUT&RUN data show that ALKBH5 can bind to DNA in Hepa1-6 cells.

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STAT5 occupancy profiling in GSCs (CUT&Tag)

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scNanoSeq-CUT&Tag: a long-read single-cell CUT&Tag sequencing method for efficient chromatin modification profiling within individual cells

Project description:We developed scNanoSeq-CUT&Tag, a streamlined method by adapting a modified CUT&Tag protocol to Oxford Nanopore sequencing platform for efficient chromatin modification profiling at single-cell resolution. We firstly tested the performance of scNanoSeq-CUT&Tag on six human cell lines: K562, 293T, GM12878, HG002, H9, HFF1 and adult mouse blood cells, it showed that scNanoSeq-CUT&Tag can accurately distinguish different cell types in vitro and in vivo. Moreover, scNanoSeq-CUT&Tag enables to effectively map the allele-specific epigenomic modifications in the human genome andallows to analyze co-occupancy of histone modifications. Taking advantage of long-read sequencing,scNanoSeq-CUT&Tag can sensitively detect epigenomic state of repetitive elements. In addition, by applying scNanoSeq-CUT&Tag to testicular cells of adult mouse B6D2F1, we demonstrated that scNanoSeq-CUT&Tag maps dynamic epigenetic state changes during mouse spermatogenesis. Finally, we exploited the epigenetic changes of human leukemia cell line K562 during DNA demethylation, it showed that NanoSeq-CUT&Tag can capture H3K27ac signals changes along DNA demethylation. Overall, we prove that scNanoSeq-CUT&Tag is a valuable tool for efficiently probing chromatin state changes within individual cells.

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Investigate Drl and Blf genome-wide occupancy in zebrafish hematopoietic progenitors with CUT&Tag

Project description:Perturbations in embryonic hematopoiesis may result in illness ranges from fetal anemia to leukemias. We found hematopoiesis in zebrafish embryo are synergistically regulated by blf and drl cluster genes. To investigate the blf and drl cluster regulatory network and mechanism of action, we used CUT&Tag technology to investigate their genome-wide occupancy in haematopoietic progenitors.

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ALKBH5 modulates endoderm fate diversion from hESCs through the ALKBH5-GATA6-WNT/β-catenin regulatory axis [CUT&Tag]

Project description:N6-methyladenonsine (m6A) modification locates ubiquitously in mammalian mRNA, and profoundly impacts various physiological processes and pathogenesis. However, the precise involvement of m6A in early endoderm development has yet to be fully elucidated. Here, we reported that depletion of the m6A demethylase ALKBH5 in human embryonic stem cells (hESCs) severely impaired definitive endoderm (DE) differentiation. Within this process, ALKBH5-/- hESCs failed to undergo the primitive streak (PS) intermediate transition, which is considered as a prelude to endoderm specification. Mechanistically, we demonstrated that ALKBH5 deficiency induced m6A hypermethylation around the 3’ untranslated region (3’UTR) of GATA6 transcripts and destabilized GATA6 mRNA in a YTHDF2-dependent manner. Moreover, dysregulation of GATA6 expression ablated its occupancy with critical regulators of Wnt/β-catenin signaling pathway, thereby disrupting the signaling logic underlying DE formation. Overall, our findings unveil a mechanism whereby the ALKBH5-GATA6-WNT/β-catenin axis modulates human in vitro DE induction, and present novel insights on m6A modification in early embryonic development.

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