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Base editing in bovine embryos reveals a species-specific role of SOX2 in regulation of pluripotency

The emergence of the first three lineages during development is orchestrated by a network of transcription factors, which are best characterized in mice. However, the role and regulation of these factors are not completely conserved in other mammals, including human and cattle. Here, we establish a...

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Autores principales: Luo, Lei, Shi, Yan, Wang, Huanan, Wang, Zizengchen, Dang, Yanna, Li, Shuang, Wang, Shaohua, Zhang, Kun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9286228/
https://www.ncbi.nlm.nih.gov/pubmed/35788719
http://dx.doi.org/10.1371/journal.pgen.1010307
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author Luo, Lei
Shi, Yan
Wang, Huanan
Wang, Zizengchen
Dang, Yanna
Li, Shuang
Wang, Shaohua
Zhang, Kun
author_facet Luo, Lei
Shi, Yan
Wang, Huanan
Wang, Zizengchen
Dang, Yanna
Li, Shuang
Wang, Shaohua
Zhang, Kun
author_sort Luo, Lei
collection PubMed
description The emergence of the first three lineages during development is orchestrated by a network of transcription factors, which are best characterized in mice. However, the role and regulation of these factors are not completely conserved in other mammals, including human and cattle. Here, we establish a gene inactivation system with a robust efficiency by introducing premature codon with cytosine base editors in bovine early embryos. By using this approach, we have determined the functional consequences of three critical lineage-specific genes (SOX2, OCT4 and CDX2) in bovine embryos. In particular, SOX2 knockout results in a failure of the establishment of pluripotency in blastocysts. Indeed, OCT4 level is significantly reduced and NANOG barely detectable. Furthermore, the formation of primitive endoderm is compromised with few SOX17 positive cells. RNA-seq analysis of single blastocysts (day 7.5) reveals dysregulation of 2074 genes, among which 90% are up-regulated in SOX2-null blastocysts. Intriguingly, more than a dozen lineage-specific genes, including OCT4 and NANOG, are down-regulated. Moreover, SOX2 level is sustained in the trophectoderm in absence of CDX2. However, OCT4 knockout does not affect the expression of SOX2. Overall, we propose that SOX2 is indispensable for OCT4 and NANOG expression and CDX2 represses the expression of SOX2 in the trophectoderm in cattle, which are all in sharp contrast with results in mice.
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spelling pubmed-92862282022-07-16 Base editing in bovine embryos reveals a species-specific role of SOX2 in regulation of pluripotency Luo, Lei Shi, Yan Wang, Huanan Wang, Zizengchen Dang, Yanna Li, Shuang Wang, Shaohua Zhang, Kun PLoS Genet Research Article The emergence of the first three lineages during development is orchestrated by a network of transcription factors, which are best characterized in mice. However, the role and regulation of these factors are not completely conserved in other mammals, including human and cattle. Here, we establish a gene inactivation system with a robust efficiency by introducing premature codon with cytosine base editors in bovine early embryos. By using this approach, we have determined the functional consequences of three critical lineage-specific genes (SOX2, OCT4 and CDX2) in bovine embryos. In particular, SOX2 knockout results in a failure of the establishment of pluripotency in blastocysts. Indeed, OCT4 level is significantly reduced and NANOG barely detectable. Furthermore, the formation of primitive endoderm is compromised with few SOX17 positive cells. RNA-seq analysis of single blastocysts (day 7.5) reveals dysregulation of 2074 genes, among which 90% are up-regulated in SOX2-null blastocysts. Intriguingly, more than a dozen lineage-specific genes, including OCT4 and NANOG, are down-regulated. Moreover, SOX2 level is sustained in the trophectoderm in absence of CDX2. However, OCT4 knockout does not affect the expression of SOX2. Overall, we propose that SOX2 is indispensable for OCT4 and NANOG expression and CDX2 represses the expression of SOX2 in the trophectoderm in cattle, which are all in sharp contrast with results in mice. Public Library of Science 2022-07-05 /pmc/articles/PMC9286228/ /pubmed/35788719 http://dx.doi.org/10.1371/journal.pgen.1010307 Text en © 2022 Luo et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Luo, Lei
Shi, Yan
Wang, Huanan
Wang, Zizengchen
Dang, Yanna
Li, Shuang
Wang, Shaohua
Zhang, Kun
Base editing in bovine embryos reveals a species-specific role of SOX2 in regulation of pluripotency
title Base editing in bovine embryos reveals a species-specific role of SOX2 in regulation of pluripotency
title_full Base editing in bovine embryos reveals a species-specific role of SOX2 in regulation of pluripotency
title_fullStr Base editing in bovine embryos reveals a species-specific role of SOX2 in regulation of pluripotency
title_full_unstemmed Base editing in bovine embryos reveals a species-specific role of SOX2 in regulation of pluripotency
title_short Base editing in bovine embryos reveals a species-specific role of SOX2 in regulation of pluripotency
title_sort base editing in bovine embryos reveals a species-specific role of sox2 in regulation of pluripotency
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9286228/
https://www.ncbi.nlm.nih.gov/pubmed/35788719
http://dx.doi.org/10.1371/journal.pgen.1010307
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