Development and application of haploid embryonic stem cells

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Abstract

Haploid cells are a kind of cells with only one set of chromosomes. Compared with traditional diploid cells, haploid cells have unique advantages in gene screening and drug-targeted therapy, due to their phenotype being equal to the genotype. Embryonic stem cells are a kind of cells with strong differentiation potential that can differentiate into various types of cells under specific conditions in vitro. Therefore, haploid embryonic stem cells have the characteristics of both haploid cells and embryonic stem cells, which makes them have significant advantages in many aspects, such as reproductive developmental mechanism research, genetic screening, and drug-targeted therapy. Consequently, establishing haploid embryonic stem cell lines is of great significance. This paper reviews the progress of haploid embryonic stem cell research and briefly discusses the applications of haploid embryonic stem cells.

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The Xist lncRNA directly interacts with SHARP to silence transcription through HDAC3

Colleen A McHugh, Chun-Kan Chen, Amy Chow … (2015)

Many long non-coding RNAs (lncRNAs) affect gene expression 1 , but the mechanisms by which they act are still largely unknown 2 . One of the best-studied lncRNAs is Xist, which is required for transcriptional silencing of one X-chromosome during development in female mammals 3,4 . Despite extensive efforts to define the mechanism of Xist-mediated transcriptional silencing, we still do not know any proteins required for this role 3 . The main challenge is that there are currently no methods to comprehensively define the proteins that directly interact with a lncRNA in the cell 5 . Here we develop a method to purify a lncRNA and identify its direct interacting proteins using quantitative mass spectrometry. We identify 10 proteins that specifically associate with Xist, three of these proteins – SHARP, SAF-A, and LBR – are required for Xist-mediated transcriptional silencing. We show that SHARP, which interacts with the SMRT co-repressor 6 that activates HDAC3 7 , is not only essential for silencing, but is also required for the exclusion of RNA Polymerase II (PolII) from the inactive X. Both SMRT and HDAC3 are also required for silencing and PolII exclusion. In addition to silencing transcription, SHARP and HDAC3 are required for Xist-mediated recruitment of the polycomb repressive complex 2 (PRC2) across the X-chromosome. Our results suggest that Xist silences transcription by directly interacting with SHARP, recruiting SMRT, activating HDAC3, and deacetylating histones to exclude PolII across the X-chromosome.

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Integration of quanta in cerebellar granule cells during sensory processing.

Paul Chadderton, Troy W. Margrie, Michael Häusser (2004)

To understand the computations performed by the input layers of cortical structures, it is essential to determine the relationship between sensory-evoked synaptic input and the resulting pattern of output spikes. In the cerebellum, granule cells constitute the input layer, translating mossy fibre signals into parallel fibre input to Purkinje cells. Until now, their small size and dense packing have precluded recordings from individual granule cells in vivo. Here we use whole-cell patch-clamp recordings to show the relationship between mossy fibre synaptic currents evoked by somatosensory stimulation and the resulting granule cell output patterns. Granule cells exhibited a low ongoing firing rate, due in part to dampening of excitability by a tonic inhibitory conductance mediated by GABA(A) (gamma-aminobutyric acid type A) receptors. Sensory stimulation produced bursts of mossy fibre excitatory postsynaptic currents (EPSCs) that summate to trigger bursts of spikes. Notably, these spike bursts were evoked by only a few quantal EPSCs, and yet spontaneous mossy fibre inputs triggered spikes only when inhibition was reduced. Our results reveal that the input layer of the cerebellum balances exquisite sensitivity with a high signal-to-noise ratio. Granule cell bursts are optimally suited to trigger glutamate receptor activation and plasticity at parallel fibre synapses, providing a link between input representation and memory storage in the cerebellum.

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Forward and reverse genetics through derivation of haploid mouse embryonic stem cells.

Ulrich Elling, Jasmin Taubenschmid, Gerald Wirnsberger … (2011)

All somatic mammalian cells carry two copies of chromosomes (diploidy), whereas organisms with a single copy of their genome, such as yeast, provide a basis for recessive genetics. Here we report the generation of haploid mouse ESC lines from parthenogenetic embryos. These cells carry 20 chromosomes, express stem cell markers, and develop into all germ layers in vitro and in vivo. We also developed a reversible mutagenesis protocol that allows saturated genetic recessive screens and results in homozygous alleles. This system allowed us to generate a knockout cell line for the microRNA processing enzyme Drosha. In a forward genetic screen, we identified Gpr107 as a molecule essential for killing by ricin, a toxin being used as a bioweapon. Our results open the possibility of combining the power of a haploid genome with pluripotency of embryonic stem cells to uncover fundamental biological processes in defined cell types at a genomic scale. Copyright © 2011 Elsevier Inc. All rights reserved.

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Author and article information

Contributors

Hai-Song Wang:

ORCID: http://orcid.org/0000-0002-6342-8042

hswang813@zzu.edu.cn

Yi-Hong Guo: 13613863710@163.com

Journal

Journal ID (nlm-ta): Stem Cell Res Ther

Journal ID (iso-abbrev): Stem Cell Res Ther

Title: Stem Cell Research & Therapy

Publisher: BioMed Central (London )

ISSN (Electronic): 1757-6512

Publication date (Electronic): 23 April 2024

Publication date PMC-release: 23 April 2024

Publication date Collection: 2024

Volume: 15

Electronic Location Identifier: 116

Affiliations

GRID grid.412633.1, ISNI 0000 0004 1799 0733, Center for Reproductive Medicine, , The First Affiliated Hospital of Zhengzhou University, Zhengzhou University, ; No. 40 Daxue Road, 450052 Zhengzhou, Henan Province China

Author information

Hai-Song Wang http://orcid.org/0000-0002-6342-8042

Article

Publisher ID: 3727

DOI: 10.1186/s13287-024-03727-y

PMC ID: 11040874

PubMed ID: 38654389

SO-VID: bb380cb9-7845-4f9c-887d-e7d003a32683

License:

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

History

Date received : 7 December 2023

Date accepted : 10 April 2024

Funding

Funded by: FundRef http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;

Award ID: 81901476

Award Recipient : Hai-Song Wang

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ScienceOpen disciplines: Molecular medicine

Keywords: haploid embryonic stem cells,genetic screening,x chromosome inactivation,haploid diploidization,gamete substitution

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ScienceOpen disciplines: Molecular medicine

Keywords: haploid embryonic stem cells, genetic screening, x chromosome inactivation, haploid diploidization, gamete substitution

Development and application of haploid embryonic stem cells

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Stress signalling in stem cells

Most cited references 53

The Xist lncRNA directly interacts with SHARP to silence transcription through HDAC3

Integration of quanta in cerebellar granule cells during sensory processing.

Forward and reverse genetics through derivation of haploid mouse embryonic stem cells.

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