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Small RNA expression analysis in histone replacement-completed sperm

ABSTRACT: Small RNA expression analysis in histone replacement-completed sperm

PROVIDER: PRJNA507970 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8271760_1.fastq.gz	Fastqsanger.gz
	SRR8271760_2.fastq.gz	Fastqsanger.gz
	SRR8271770_1.fastq.gz	Fastqsanger.gz
	SRR8271770_2.fastq.gz	Fastqsanger.gz
	SRR8271780_1.fastq.gz	Fastqsanger.gz

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Mapping of histone-binding sites in histone replacement-completed spermatozoa

Project description:Mapping of histone-binding sites in histone replacement-completed spermatozoa

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Small RNA expression analysis in histone replacement-completed sperm

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Project description:Majority of histones are replaced by protamine during spermatogenesis, but small amount of histones are retained in mammalian spermatozoa. Since nucleosomes in spermatozoa influence epigenetic inheritance, it is important how histones are distributed on sperm genome. We have found that the swim-up sperm used in many studies contains about 10% of immature sperm which do not yet complete the histone-to-protamine replacement. We have developed the novel method to purify the histone replacement-completed sperm (HRCS), and to completely solubilize nucleosomes from cross-linked HRCS without MNase digestion. Here, we analyzed H3 binding profiles in total sperm and HRCS by Chromatin IP – sequencing (ChIP-seq) analysis, and DNA methylation profiles in swim-up sperm and HRCS by target bisulfite sequencing (TGSB).

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Project description:The majority of histones are replaced by protamines during spermatogenesis, but small amounts are retained in mammalian spermatozoa. Since nucleosomes in spermatozoa influence epigenetic inheritance, it is important to know how histones are distributed in the sperm genome. Conflicting data, which may result from different conditions used for micrococcal nuclease (MNase) digestion, have been reported: retention of nucleosomes at either gene promoter regions or within distal gene-poor regions. Here, we find that the swim-up sperm used in many studies contain about 10% population of sperm which have not yet completed the histone-to-protamine replacement. We develop a method to purify histone replacement-completed sperm (HRCS) and to completely solubilize histones from cross-linked HRCS without MNase digestion. Our results indicate that histones are retained at specific promoter regions in HRCS. This method allows the study of epigenetic status in mature sperm.

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Mapping of histone-binding sites in histone replacement-completed spermatozoa

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