Analysis and modulation of H3K9 methylation in bovine somatic cells

• Grant number: 15/25111-5
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Effective date (Start): March 01, 2016
• Effective date (End): February 28, 2017
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Flávio Vieira Meirelles
• Grantee: Rafael Vilar Sampaio
• Supervisor: Pablo Juan Ross
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
• Research place: University of California, Davis (UC Davis), United States
• Associated to the scholarship: 15/08807-6 - H3K9 methylation as regulator of epigenetic memory on bovine nuclear reprograming., BP.PD

Abstract

Epigenetic mechanisms are responsible for many cellular functions. Amongst them, is the epigenetic memory maintained during cell division. Consolidation of this information is made by stable chemical marks as DNA methylation and histone modification. Understanding epigenetic mechanism allows the development of several studies, such as nuclear reprogramming, which is directly influenced by those marks. Thus, H3K9 methylation has been shown as the main barrier on the rodents nuclear remodeling. Hence, the main goal of this proposal is to investigate genes directly regulated by H3K9 methylation, and modulate their pattern of expression through H3K9 modifications of bovine cells. For this, two strategies were elaborated: Firstly, bovine fetal fibroblast (bff) genes regulated by H3K9 will be identify by ChIP-seq analyses using antibodies anti-H3K9me2 and H3K9me3. Then, we aim to modify the H3K9 methylation levels on bff. In order to achieve these modifications histone methyltransferases enzymes (G9a and SUV39h1) will be silenced by small interference RNA technique (siRNA). Secondly, the H3K9me2, H3K9me3, 5mC and H3K27 levels of siRNA treated bff will be analyzed by immunostaing. Therefore this proposal presents a unique opportunity to discovery groundbreaking epigenetic related events. In addition, results obtained in this project will be applied on several fields of knowledge related with human and animal health. It is noteworthy, that this BEPE will be prepare in collaboration with Dr. Pablo Ross at DAVIS University, which has extensive expertise on the epigenetic of livestock animals. Furthermore, another goal of this research internship is to capacitate and train the candidate to be able to use all these innovative technologies. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

SAMPAIO, RAFAEL VILAR; SANGALLI, JULIANO RODRIGUES; CAMARA DE BEM, TIAGO HENRIQUE; AMBRIZI, DEWISON RICARDO; DEL COLLADO, MAITE; BRIDI, ALESSANDRA; CAVALCANTE MENDES DE AVILA, ANA CLARA FAQUINELI; MACABELLI, CAROLINA HABERMANN; OLIVEIRA, LILIAN DE JESUS; DA SILVEIRA, JULIANO COELHO; et al. Catalytic inhibition of H3K9me2 writers disturbs epigenetic marks during bovine nuclear reprogramming. SCIENTIFIC REPORTS, v. 10, n. 1, JUL 13 2020. (15/08807-6, 14/22887-0, 13/08135-2, 14/50947-7, 14/21034-3, 15/25111-5, 12/50533-2, 13/07160-3)

BOGLIOTTI, YANINA SOLEDAD; WU, JUN; VILARINO, MARCELA; OKAMURA, DAIJI; SOTO, DELIA ALBA; ZHONG, CUIQING; SAKURAI, MASAHIRO; SAMPAIO, RAFAEL VILAR; SUZUKI, KEIICHIRO; BELMONTE, JUAN CARLOS IZPISUA; et al. Efficient derivation of stable primed pluripotent embryonic stem cells from bovine blastocysts. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v. 115, n. 9, p. 2090-2095, FEB 27 2018. (15/25111-5, 13/08135-2)