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Capturing totipotency in human cells through spliceosomal repression [RNA-Seq]

ABSTRACT: Supplemented with splicing inhibitor PlaB, we realize in vitro culturing of human totipotent stem cells comparable to 2-4-cellb blastomeres Poly(A) RNA-seq is performed on hESCs cultured in MYCP medium

ORGANISM(S): Homo sapiens

PROVIDER: GSE224794 | GEO | 2024/05/30

REPOSITORIES: GEO

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Capturing totipotency in human cells through spliceosomal repression [scRNA-seq]

Project description:Supplemented with splicing inhibitor PlaB, we realize in vitro culturing of human totipotent stem cells comparable to 2-4-cell blastomeres

2024-05-30 | GSE226228 | GEO

Capturing totipotency in human cells through spliceosomal repression [ATAC-seq]

Project description:Supplemented with splicing inhibitor PlaB, we realize in vitro culturing of human totipotent stem cells comparable to 2-4-cell blastomeres Assay for transposase accessible chromatin with high-throughput sequencing in hESC sand hTBLCs

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Capturing totipotency in human cells through spliceosomal repression [ChIP-seq]

Project description:Supplemented with splicing inhibitor PlaB, we realize in vitro culturing of human totipotent stem cells comparable to 2-4-cell blastomeres Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for h3k4me3 and h3k27me3 in hESC sand hTBLCs

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Mouse totipotent stem cells captured and maintained through spliceosomal repression

Project description:Since the establishment of the first embryonic stem cells (ESCs), in vitro culture of totipotent cells functionally and molecularly comparable to in vivo blastomeres with embryonic and extraembryonic developmental potency is unviable. Spliceosomes are responsible for mRNA splicing and maturation. Here, we report that spliceosomal repression in mouse ESCs drives pluripotent-to-totipotent state transition. Using the splicing inhibitor Pladienolide B, we realize in vitro culturing of totipotent ESCs comparable to 2- and 4-cell blastomeres at molecular levels for long-time passages, which are therefore termed as totipotent blastomere-like cells (TBLCs). Mouse chimeric assays combined with single-cell RNA-seq technology demonstrate that TBLCs own a robust bidirectional development capability to generate multiple embryonic and extraembryonic cell lineages. Mechanically, spliceosomal repression causes widespread splicing inhibition of pluripotent genes, whereas the totipotent genes featured with few short introns are efficiently spliced and transcriptionally activated. Our study provides a principle for capturing and maintenance of totipotent stem cells.

2021-05-15 | GSE168728 | GEO

Capturing totipotency in human cells through spliceosomal repression

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

2024-05-30 | GSE226231 | GEO

Capturing totipotency in human cells through spliceosomal repression

Project description:Capturing totipotency in human cells through spliceosomal repression

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Totipotency of mouse zygotes extends to single blastomeres of 4-cell stage embryos

Project description:In multicellular organisms, heterogametes of oocytes and sperms are fertilized and resulting zygotes give rise to new individuals. The ability of zygotes that produce a fully formed individual from single cell when placed in a supportive environment is defined as totipotency. Given that totipotent cells are the source of all multicellular organisms, better understanding of totipotency has a profound effect on not only biology but also our society. However, the exact distribution of totipotent cells in mammals remains elusive, although zygotes and single blastomeres at 2-cell stage embryos has been thought to be only mouse cells to be totipotent. We now show that a single blastomere isolated from 2- and 4-cell stage embryos gives rise to a fertile adult individual when it placed in a uterus, although isolation of blastomeres at these stage results in the disturbance of transcriptome in single blastomere derived embryos. Single blastomeres from 8-cell and morula stage embryos that were separately cultured in vitro exhibited severe defects in the formation of epiblast and primitive endoderm in the inner cell mass and the development to blastocysts, respectively. Our results thus indicate that totipotency of mouse zygotes extends to single blastomeres of 4-cell stage embryos.

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Defining totipotency using criteria of increasing stringency

Project description:Totipotency is the ability of a single cell to give rise to all the differentiated cells that build the conceptus, yet how to capture this property in vitro remains incompletely understood. Defining totipotency relies upon a variety of assays of variable stringency. Here we describe criteria to define totipotency. We illustrate how distinct criteria of increasing stringency can be used to judge totipotency by evaluating candidate totipotent cell types in the mouse, including early blastomeres and expanded or extended pluripotent stem cells. Our data challenge the notion that expanded or extended pluripotent states harbor increased totipotent potential relative to conventional embryonic stem cells under in vivo conditions.

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Capturing totipotency in human cells through spliceosomal repression [scRNA-seq]

Project description:Capturing totipotency in human cells through spliceosomal repression [scRNA-seq]

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Project description:Capturing totipotency in human cells through spliceosomal repression [ATAC-seq]

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