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植入后单倍体胚胎干细胞中的高通量筛选揭示 Hs3st3b1 可作为重编程的调节剂。

High-throughput screening in postimplantation haploid epiblast stem cells reveals Hs3st3b1 as a modulator for reprogramming.

机构信息

State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin, People's Republic of China.

Department of Obstetrics, Tianjin First Central Hospital, Nankai University, Tianjin, People's Republic of China.

出版信息

Stem Cells Transl Med. 2021 May;10(5):743-755. doi: 10.1002/sctm.20-0468. Epub 2021 Jan 29.

DOI:10.1002/sctm.20-0468
PMID:33511777
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8046116/
Abstract

Epiblast stem cells (EpiSCs) derived from postimplantation epiblast are pluripotent stem cells, epigenetically distinct from embryonic stem cells (ESCs), which are widely used in reprogramming studies. Recent achieved haploid cell lines in mammalian species open a new era for high-throughput genetic screening, due to their homozygous phenotypes. Here, we report the generation of mouse haploid EpiSCs (haEpiSCs) from postimplantation chimeric embryos at embryonic day 6.5 (E6.5). These cells maintain one set of chromosomes, express EpiSC-specific genes, and have potentials to differentiate into three germ layers. We also develop a massive mutagenesis protocol with haEpiSCs, and subsequently perform reprogramming selection using this genome-wide mutation library. Multiple modules related to various pathways are implicated. The validation experiments prove that knockout of Hst3st3b1 (one of the candidates) can promote reprogramming of EpiSCs to the ground state efficiently. Our results open the feasibility of utilizing haEpiSCs to elucidate fundamental biological processes including cell fate alternations.

摘要

内细胞团干细胞（EpiSCs）来源于着床后的上胚层，是多能干细胞，在表观遗传学上有别于胚胎干细胞（ESCs），后者被广泛用于重编程研究。最近在哺乳动物中获得的单倍体细胞系由于其纯合表型，为高通量遗传筛选开辟了一个新时代。在这里，我们报告了从胚胎 6.5 天（E6.5）的嵌合胚胎中生成小鼠单倍体 EpiSCs（haEpiSCs）。这些细胞保持一组染色体，表达 EpiSC 特异性基因，并具有分化为三个胚层的潜力。我们还开发了一种大规模的突变生成方案，并随后使用这个全基因组突变文库进行重编程筛选。多个与各种途径相关的模块被涉及。验证实验证明，敲除 Hst3st3b1（候选者之一）可以有效地促进 EpiSCs 向基础状态的重编程。我们的结果为利用 haEpiSCs 来阐明包括细胞命运转换在内的基本生物学过程开辟了可行性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/1b52cd5d4f8d/SCT3-10-743-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/da8b92b9dddd/SCT3-10-743-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/cd568f454afe/SCT3-10-743-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/4543e44abc73/SCT3-10-743-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/6abfd7eb125f/SCT3-10-743-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/1b52cd5d4f8d/SCT3-10-743-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/da8b92b9dddd/SCT3-10-743-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/cd568f454afe/SCT3-10-743-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/4543e44abc73/SCT3-10-743-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/6abfd7eb125f/SCT3-10-743-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ed8/8046116/1b52cd5d4f8d/SCT3-10-743-g001.jpg

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