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NANOG is required to form the epiblast and maintain pluripotency in the bovine embryo
Author(s) -
Ortega M. Sofia,
Kelleher Andrew M.,
O'Neil Eleanore,
Benne Joshua,
Cecil Raissa,
Spencer Thomas E.
Publication year - 2020
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.23304
Subject(s) - epiblast , homeobox protein nanog , biology , nanog homeobox protein , sox2 , inner cell mass , rex1 , microbiology and biotechnology , embryo , embryonic stem cell , blastocyst , gastrulation , genetics , induced pluripotent stem cell , embryogenesis , gene
During preimplantation development, the embryo undergoes two consecutive lineages specifications. The first cell fate decision determines which cells give rise to the trophectoderm (TE) and the inner cell mass (ICM). Subsequently, the ICM differentiates into hypoblast and epiblast, the latter giving rise to the embryo proper. The transcription factors that govern these cell fate decisions have been extensively studied in the mouse, but are still poorly understood in other mammalian species. In the present study, the role of NANOG in the formation of the epiblast and maintenance of pluripotency in the bovine embryo was investigated. Using a CRISPR‐Cas9 approach, guide RNAs were designed to target exon 2, resulting in a functional deletion of bovine NANOG at the zygote stage. Disruption of NANOG resulted in the embryos that form a blastocoel and an ICM composed of hypoblast cells. Furthermore, NANOG ‐null embryos showed lower expression of epiblast cell markers SOX2 and HA2AFZ , and hypoblast marker GATA6 ; without affecting the expression of TE markers CDX2 and KRT8 . Results indicate that NANOG , has no apparent role in segregation or maintenance of the TE, but it is required to derive and maintain the pluripotent epiblast and during the second lineage commitment in the bovine embryo.