Valoración del papel de PPARγ durante el desarrollo embrionario bovino mediante la técnica CRISPR/Cas

Argüeso Gómez, Matilde (2020). Valoración del papel de PPARγ durante el desarrollo embrionario bovino mediante la técnica CRISPR/Cas. Proyecto Fin de Carrera / Trabajo Fin de Grado, E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM), Madrid, España.

Description

Title: Valoración del papel de PPARγ durante el desarrollo embrionario bovino mediante la técnica CRISPR/Cas
Author/s:
  • Argüeso Gómez, Matilde
Contributor/s:
  • García Rebollar, Pilar
Item Type: Final Project
Degree: Grado en Biotecnología
Date: July 2020
Subjects:
Faculty: E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM)
Department: Producción Agraria
Creative Commons Licenses: Recognition - No derivative works - Non commercial

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Abstract

Prostaglandins (PGs) and other prostanoids are important regulators of reproductive processes including the maternal-embryonic signaling, the establishment of pregnancy and early embryonic development. Prostaglandin signaling is widely mediated by PPAR receptors (peroxisome proliferator-activated receptors), being PPARγ an isotype of these receptors expressed by the embryo before and during conceptus elongation, which function has not been fully elucidated. The objective of this work has been to analyze the function of PPARγ-mediated signaling during early embryonic development by generating PPARγ KO bovine embryos by in vitro oocyte maturation, fertilization and culture. Oocytes were obtained from ovaries from the slaughterhouse, and following in vitro maturation, PPARγ was ablated via CRISPR/Cas microinjection (group Cas9+guide). Embryos microinjected exclusively with Cas9-coding mRNA were used as control (group Cas9). The parameters analyzed were the developmental rates to the blastocyst stage and blastocyst cell number determined by nuclear staining with DAPI followed by structured illumination microscopy. To determine embryo genotype (i.e. WT, Hz or KO), genotyping was performed by clonal sequencing. The comparison of these three groups enabled to investigate if PPARG fulfills essential functions during bovine preimplantation embryonic development. In conclusion, our results show that PPARG does not have a fundamental function in this period of embryonic development since the developmental rates to blastocyst were similar in both experimental groups (Cas9+guide and Cas9) and the number of cells in the blastocysts did not present significant differences between the KO, WT and Hz embryos.

More information

Item ID: 66711
DC Identifier: https://oa.upm.es/66711/
OAI Identifier: oai:oa.upm.es:66711
Deposited by: Biblioteca ETSI Agrónomos
Deposited on: 13 Apr 2021 10:38
Last Modified: 13 Jun 2021 22:30
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