{"title":"LncRNA <i>XLOC-040580</i> targeted by <i>TPRA1</i> coordinate zygotic genome activation during porcine embryonic development.","authors":"Mengxin Liu, Enhong Li, Haiyuan Mu, Zimo Zhao, Xinze Chen, Jie Gao, Dengfeng Gao, Zhiyu Liu, Jianyong Han, Liang Zhong, Suying Cao","doi":"10.1177/09636897251332527","DOIUrl":null,"url":null,"abstract":"<p><p>Long noncoding RNAs (lncRNAs) are crucial in porcine preimplantation embryonic development, yet their regulatory role during zygote genome activation (ZGA) is poorly understood. We analyzed transcriptome data from porcine fetal fibroblasts (PEF), induced pluripotent stem cells (iPS), and preimplantation embryos, identifying ZGA-specific lncRNAs like <i>XLOC-040580</i>, and further predicted its potentially interacting genes <i>TPRA1</i> and <i>BCL2L1</i> via co-expression network. <i>XLOC-040580</i> was knocked down by siRNA microinjection and the expression of ZGA-related genes was detected by qRT-PCR. After microinjecting siRNA targeting <i>TPRA1</i> and <i>BCL2L1</i> at the one-cell stage, we counted the blastocyst development rate. The blastocyst development rate was consistent with the results from si-XLOC-040580 after si-<i>TPRA1</i>. Through dual-luciferase reporter assays, we found that <i>XLOC-040580</i> was a downstream target of <i>TPRA1</i>. To further elucidate the mechanism of <i>XLOC-040580</i>, Single-cell mRNA sequencing after <i>XLOC-040580</i> knockdown revealed its regulatory network involved in embryonic developmental defects. Transcriptome analysis revealed that <i>XLOC-040580</i> was specifically expressed during zygote activation. Knockdown of <i>XLOC-040580</i> decreased the blastocyst development rate and reduced both the total blastocyst cell number and TE cell number. <i>TPRA1</i> and <i>BCL2L1 were</i> specifically co-expressed with <i>XLOC-040580</i> during ZGA stage, and <i>TPRA1</i> could interact with the promoter region of <i>XLOC-040580</i> and regulate its expression. Knockdown of <i>TPRA1</i> or <i>XLOC-040580</i> blocked porcine embryonic development by affecting the expression of ZGA-related genes. We found and validated that lncRNA <i>XLOC-040580</i> played a key role in the ZGA process, which was regulated by <i>TPRA1</i>. These results implied that the functional axis of <i>TPRA1</i>-<i>XLOC-040580</i>-downstream genes involved in ZGA-related functions also coordinated early embryonic development in porcine.</p>","PeriodicalId":9721,"journal":{"name":"Cell Transplantation","volume":"34 ","pages":"9636897251332527"},"PeriodicalIF":3.2000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12035016/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cell Transplantation","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1177/09636897251332527","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/4/17 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"CELL & TISSUE ENGINEERING","Score":null,"Total":0}
引用次数: 0
Abstract
Long noncoding RNAs (lncRNAs) are crucial in porcine preimplantation embryonic development, yet their regulatory role during zygote genome activation (ZGA) is poorly understood. We analyzed transcriptome data from porcine fetal fibroblasts (PEF), induced pluripotent stem cells (iPS), and preimplantation embryos, identifying ZGA-specific lncRNAs like XLOC-040580, and further predicted its potentially interacting genes TPRA1 and BCL2L1 via co-expression network. XLOC-040580 was knocked down by siRNA microinjection and the expression of ZGA-related genes was detected by qRT-PCR. After microinjecting siRNA targeting TPRA1 and BCL2L1 at the one-cell stage, we counted the blastocyst development rate. The blastocyst development rate was consistent with the results from si-XLOC-040580 after si-TPRA1. Through dual-luciferase reporter assays, we found that XLOC-040580 was a downstream target of TPRA1. To further elucidate the mechanism of XLOC-040580, Single-cell mRNA sequencing after XLOC-040580 knockdown revealed its regulatory network involved in embryonic developmental defects. Transcriptome analysis revealed that XLOC-040580 was specifically expressed during zygote activation. Knockdown of XLOC-040580 decreased the blastocyst development rate and reduced both the total blastocyst cell number and TE cell number. TPRA1 and BCL2L1 were specifically co-expressed with XLOC-040580 during ZGA stage, and TPRA1 could interact with the promoter region of XLOC-040580 and regulate its expression. Knockdown of TPRA1 or XLOC-040580 blocked porcine embryonic development by affecting the expression of ZGA-related genes. We found and validated that lncRNA XLOC-040580 played a key role in the ZGA process, which was regulated by TPRA1. These results implied that the functional axis of TPRA1-XLOC-040580-downstream genes involved in ZGA-related functions also coordinated early embryonic development in porcine.
期刊介绍：
Cell Transplantation, The Regenerative Medicine Journal is an open access, peer reviewed journal that is published 12 times annually. Cell Transplantation is a multi-disciplinary forum for publication of articles on cell transplantation and its applications to human diseases. Articles focus on a myriad of topics including the physiological, medical, pre-clinical, tissue engineering, stem cell, and device-oriented aspects of the nervous, endocrine, cardiovascular, and endothelial systems, as well as genetically engineered cells. Cell Transplantation also reports on relevant technological advances, clinical studies, and regulatory considerations related to the implantation of cells into the body in order to provide complete coverage of the field.
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