CRISPR Screening Reveals a Novel Role for FOXH1 in Regulating Pluripotency of Porcine Embryonic Stem Cells

IF 14.1 1区 材料科学 Q1 CHEMISTRY, MULTIDISCIPLINARY
Peng Su, Linhui Wu, Delong Li, Wenting Song, Dagang Tao, Liang Liu, Qi Wang, Manxin Gao, Tian Xu, Xin Liu, Shengsong Xie, Xia Zhang, Jilong Zhou, Yi-Liang Miao
{"title":"CRISPR Screening Reveals a Novel Role for FOXH1 in Regulating Pluripotency of Porcine Embryonic Stem Cells","authors":"Peng Su,&nbsp;Linhui Wu,&nbsp;Delong Li,&nbsp;Wenting Song,&nbsp;Dagang Tao,&nbsp;Liang Liu,&nbsp;Qi Wang,&nbsp;Manxin Gao,&nbsp;Tian Xu,&nbsp;Xin Liu,&nbsp;Shengsong Xie,&nbsp;Xia Zhang,&nbsp;Jilong Zhou,&nbsp;Yi-Liang Miao","doi":"10.1002/advs.202509495","DOIUrl":null,"url":null,"abstract":"<p>Porcine extended potential stem cells (pEPSCs), which exhibit both self-renewal and pluripotency, are promising for application in both agricultural biotechnology and regenerative medicine. However, the molecular mechanisms governing these two interconnected properties remain elusive. Here, two types of CRISPR-Cas9 screenings are conducted in pEPSCs. This fitness screening identified several genes essential for cell viability, including <i>PRMT1</i>, <i>MYBL2</i>, and <i>NASP</i>. Concurrently, FACS-based screenings revealed genes crucial for pluripotency, such as <i>SOX2</i>, <i>ZFP42</i>, and <i>FOXH1</i>. Notably, it is demonstrated that FOXH1 is required for maintaining pluripotency in pEPSCs, which complements the understanding of its role in mesendoderm specification. pEPSCs lacking FOXH1 exhibited a flatter and more dispersed clonal morphology, accompanied by downregulation of pluripotency genes and upregulation of lineage-specific genes. Additionally, <i>FOXH1</i> knockdown significantly impaired blastocyst formation during early pig embryogenesis. Functionally, the dual role of FOXH1 in pluripotency maintenance and cell differentiation is validated: FOXH1 preserves pluripotency by enhancing chromatin accessibility at pluripotency gene loci, while also influencing lineage specification through H3K4me3 modification at developmental related genes. Thus, these findings uncover a novel role of FOXH1 involved in the core regulatory network that orchestrates gene expression programs to maintain the pluripotency state of pEPSCs and provide valuable insights into categorizing gene function.</p>","PeriodicalId":117,"journal":{"name":"Advanced Science","volume":"12 34","pages":""},"PeriodicalIF":14.1000,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12442679/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Advanced Science","FirstCategoryId":"88","ListUrlMain":"https://advanced.onlinelibrary.wiley.com/doi/10.1002/advs.202509495","RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}
引用次数: 0

Abstract

Porcine extended potential stem cells (pEPSCs), which exhibit both self-renewal and pluripotency, are promising for application in both agricultural biotechnology and regenerative medicine. However, the molecular mechanisms governing these two interconnected properties remain elusive. Here, two types of CRISPR-Cas9 screenings are conducted in pEPSCs. This fitness screening identified several genes essential for cell viability, including PRMT1, MYBL2, and NASP. Concurrently, FACS-based screenings revealed genes crucial for pluripotency, such as SOX2, ZFP42, and FOXH1. Notably, it is demonstrated that FOXH1 is required for maintaining pluripotency in pEPSCs, which complements the understanding of its role in mesendoderm specification. pEPSCs lacking FOXH1 exhibited a flatter and more dispersed clonal morphology, accompanied by downregulation of pluripotency genes and upregulation of lineage-specific genes. Additionally, FOXH1 knockdown significantly impaired blastocyst formation during early pig embryogenesis. Functionally, the dual role of FOXH1 in pluripotency maintenance and cell differentiation is validated: FOXH1 preserves pluripotency by enhancing chromatin accessibility at pluripotency gene loci, while also influencing lineage specification through H3K4me3 modification at developmental related genes. Thus, these findings uncover a novel role of FOXH1 involved in the core regulatory network that orchestrates gene expression programs to maintain the pluripotency state of pEPSCs and provide valuable insights into categorizing gene function.

Abstract Image

Abstract Image

Abstract Image

Abstract Image

CRISPR筛选揭示FOXH1在调节猪胚胎干细胞多能性中的新作用
猪扩展潜能干细胞(pEPSCs)具有自我更新和多能性,在农业生物技术和再生医学领域具有广阔的应用前景。然而,控制这两种相互关联特性的分子机制仍然难以捉摸。这里,在pEPSCs中进行了两种类型的CRISPR-Cas9筛选。这种适应性筛选确定了几个对细胞活力至关重要的基因,包括PRMT1、MYBL2和NASP。同时,基于facs的筛选揭示了对多能性至关重要的基因,如SOX2、ZFP42和FOXH1。值得注意的是,研究表明FOXH1是维持pEPSCs多能性所必需的,这补充了对其在中胚层规范中的作用的理解。缺乏FOXH1的pEPSCs表现出更平坦和更分散的克隆形态,并伴有多能性基因的下调和谱系特异性基因的上调。此外,FOXH1基因敲低显著损害了猪胚胎发生早期的囊胚形成。在功能上,FOXH1在多能性维持和细胞分化中的双重作用得到了验证:FOXH1通过增强多能性基因位点的染色质可及性来保持多能性,同时也通过修饰发育相关基因的H3K4me3来影响谱系规范。因此,这些发现揭示了FOXH1参与核心调控网络的新作用,该网络协调基因表达程序以维持pEPSCs的多能性状态,并为基因功能分类提供了有价值的见解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Advanced Science
Advanced Science CHEMISTRY, MULTIDISCIPLINARYNANOSCIENCE &-NANOSCIENCE & NANOTECHNOLOGY
CiteScore
18.90
自引率
2.60%
发文量
1602
审稿时长
1.9 months
期刊介绍: Advanced Science is a prestigious open access journal that focuses on interdisciplinary research in materials science, physics, chemistry, medical and life sciences, and engineering. The journal aims to promote cutting-edge research by employing a rigorous and impartial review process. It is committed to presenting research articles with the highest quality production standards, ensuring maximum accessibility of top scientific findings. With its vibrant and innovative publication platform, Advanced Science seeks to revolutionize the dissemination and organization of scientific knowledge.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信