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Functional evaluation of NANOG gene in segregation of cell lineages in bovine embryos

Grant number: 19/22649-5
Support type:Scholarships in Brazil - Master
Effective date (Start): November 01, 2019
Effective date (End): October 31, 2021
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal Investigator:Marcelo Demarchi Goissis
Grantee:Letícia Escobar Carreiro
Home Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Associated research grant:17/09576-3 - Studies on cell differentiation mechanisms during early bovine embryo development, AP.JP

Abstract

The second event of mammalian cell differentiation is the separation between epiblast (EPI) and hypoblast or primitive endoderm (PE). This occurs shortly after segregation of the internal cell mass (ICM) and the trophectoderm (TE). It is known in mice that NANOG acts oppositely to GATA6 and FGF signaling pathway in EPI/PE segregation. Gene expression analyzes group cell populations so that NANOG and GATA6 expression are mutually exclusive. This project aims to test the hypothesis that NANOG is required for the specification of EPI in bovine embryos. CRISPR/Cas9-mediated gene deletion of NANOG in in vitro produced zygotes is intended. Embryos will then be cultured for blastocyst formation rate, cell distribution, gene and protein expression. It is expected that EPI formation will be inhibited, which will lead to an ICM containing only PE cells. Thus this project would contribute to the understanding of the EPI/PE segregation mechanisms in the bovine species. (AU)