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RNA m6A methylome in mouse oocytes and preimplantation embryos

ABSTRACT: RNA m6A methylome in mouse oocytes and preimplantation embryos

PROVIDER: PRJNA791403 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17299200_1.fastq.gz	Fastqsanger.gz
	SRR17299200_2.fastq.gz	Fastqsanger.gz
	SRR17299201_1.fastq.gz	Fastqsanger.gz
	SRR17299201_2.fastq.gz	Fastqsanger.gz
	SRR17299202_1.fastq.gz	Fastqsanger.gz

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RNA m6A methylome in mouse oocytes and preimplantation embryos

Project description:N6-methyladenosine (m6A), the most prevalent internal modification in eukaryotic messenger RNA, plays diverse regulatory roles in many biological processes and is involved in a variety of physiological behaviors. Here we present the m6A methylome maps of mouse oocytes and preimplantation embryos by a low-input approach picoMeRIP-seq. We revealed that m6A was largely and dynamically deposited on maternal and zygotic RNAs during maternal-to-zygotic transition (MZT), especially on those encoding the factors essential for cell fate determination. m6A frequently marked the maternally-derived RNAs to be degraded and the genes that were activated during zygotic genome activation. Moreover, the RNAs derived from retrotransposons, such as MTA and MERVL, were heavily occupied by m6A. Collectively, our results provide a foundation for future studies exploring the regulatory roles of m6A in mammalian early embryo development.

2023-02-10 | GSE192440 | GEO

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Single Cell DNA Methylome Sequencing of Human Preimplantation Embryos

Project description:Although the DNA methylome of human early embryos has been analyzed, some of the key features have not been addressed to date. Here, we performed single-cell DNA methylome sequencing for human preimplantation embryos and found that tens of thousands of genomic loci exhibited de novo DNA methylation. This finding indicates that genome-wide DNA methylation reprogramming during preimplantation development is a dynamic balance between strong global demethylation and significant focused re-methylation. Furthermore, the demethylation of the paternal genome is much faster and thorough than that of the maternal genome. From the 2-cell to post-implantation stage, methylation of the paternal genome is consistently lower than that on the maternal genome. We also showed that the genetic lineage of the early blastomeres could be traced by DNA methylation analysis. Our work paves the way for deciphering the secrets of DNA methylation reprogramming in human early embryos.

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RNA m6A methylome from zebrafish zygotes, mouse liver polyA-selected RNAs, mouse embryonic stem cells, spike-in RNA controls, and single mouse oocytes and embryos

Project description:N6-methyladenosine (m6A) is a prevalent RNA modification that has key roles in distinct biological processes, including meiosis and early embryo development. Despite advances in methodology for m6A mapping, the applicability of current methods is limited by the need for large amounts of input material. We develop a sensitive picogram scale m6A RNA immunoprecipitation and sequencing (picoMeRIP-seq) method, also suitable for single-cell (scMeRIP-seq). We validate the down-scaling of the method by profiling m6A from picogram amounts of mouse liver polyA-selected RNA, mouse embryonic stem cells, and zebrafish zygotes. We provide proof-of-principle of m6A profiling in single mouse oocytes and preimplantation embryos. Our work opens a new avenue for studying m6A in single cells and scarce cell types in a transcriptome-wide manner.

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Comprehensive analysis of H3K9me2 oocytes, together with DNA methylation and expression analysis of G9a conditional KO oocytes and maternal KO preimplantation embryos

Project description:We report the first genome-wide landscape of H3K9me2 ChIP-seq profile in mouse oocytes. We also performed whole-genome bisulfite sequencing and RNA-seq analysis of G9a conditional KO oocytes and maternal KO preimplantation embryos. Our findings illuminate the functional importance of G9a in preimplantation development and, in addition, pose a question on the proposed role for H3K9me2 in protection of the maternal genome from active CG demethylation.

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Project description:Single Cell DNA Methylome Sequencing of Human Preimplantation Embryos

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Project description:Purpose: The goal of this study was to generate paired-end mRNA Seq transcriptomes of control and Kdm4a mutant oocytes, 2-cell, 4-cell and 8-cell preimplantation embryos to study the dynamics of differentially expressed genes during early development Methods: MII oocytes were isolated from wildtype and Kdm4a knockout mice. 16 MII oocytes from each genotype were washed in M2 medim after zona pellucida removal, washed once in nuclease free water and directly lysed and cDNA prepared according to the manufacturers instructions. Meanwhile, control and knockout MII oocytes were also in vitro fertilised with Kdm4a knockout sperm to produce control (+/-) and maternal mutant (-/-) embryos, and cultured in KSOM EmbryoMax medium (Sigma). 16 embryos were staged each day and harvested according to control embryos. Healthiest mutants were chosen for comparison with minimal secondary effects arising from dead/dying embryos. Similar to oocytes, individual cDNA were prepared and amplified according to manufacturers instructions in nuclease free conditions. Conclusions: Our study represents a detailed analysis of differentially expressed genes in oocytes and embryos lacking maternal and endogenous Kdm4a, with sixteen biological replicates, generated by SMARTSeq v4 paired end RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles between the samples at each developmental stage. Our results show that there is a consistent downrgulation of zygotic genome activation (ZGA) genes in the mutant 2-cell embryos. Mutant 4 and 8-cell embryos have significant upregulation of minor ZGA genes giving us robust statistical significance. We conclude that Kdm4a is required for proper gene activation during maternal-to zygotic transition in conjuction with a permissive chromatin landscape.

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Tracing the expression of circular RNAs in human preimplantation embryos

Project description:We systematically analyzed transcriptome in individual human oocytes and preimplantation embryos with a new single-cell RNA seq method we recently developed. We sequenced individual single cells or embryos at seven consecutive stages (mature oocytes, zygotes, 2-cell, 4-cell, 8-cell embryos, morulae, and blastocysts) and single hESC using single-cell RNA seq.

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