EZH2 cloning and retroviral preparation for lens primary cell culture transfection

• Grant number: 17/12112-9
• Support Opportunities: Scholarships abroad - Research Internship - Scientific Initiation
• Effective date (Start): August 03, 2017
• Effective date (End): September 04, 2017
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Vera Paschon
• Grantee: Felipe Fernandes Correia
• Supervisor: Jean Jiang
• Host Institution: Centro de Matemática, Computação e Cognição (CMCC). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil
• Research place: University of Texas Health Science Center at San Antonio (UTHSCSA), United States
• Associated to the scholarship: 15/24774-0 - Evaluation of histone deacetylases inhibition in the neuronal death spread and in the function recovery after spinal cord transection in rats, BP.IC

Abstract

Enhancer of zeste homolog 2 (EZH2) is a component of the Polycomb repressive complex (PRC) 2 with enzymatic properties. It is known to be responsible for specific gene transcriptional repress by the methylation of lysine 27 at histone H3 (H3K27me3), among other functions described more recently. Although EZH2 was well studied in the cancer field, this enzyme function is not well understood in the nervous system. Retrovirus-mediated gene transfer is a powerful molecular tool that can be used to understand gene functions. Retrovirus contains a single stranded RNA genome, which can be modified by inserting in a double stranded DNA molecule of the target cell genome. This means that retroviral vectors can be used to permanently modify the host cell nuclear genome. The aim of this project is to prepare recombinant retrovirus for the overexpression of EZH2 full-length and mutant genes in lens primary culture in order to learn the technique for further application in spinal cord cell cultures. (AU)