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Projects. Life and Matter Sciences

The acetylomes of neural differentiation

Lead Researcher:
Mario Fernández Fraga

Research Centre:
Instituto Universitario de Oncología del Principado de Asturias (IUOPA). Universidad de Oviedo.


Mario Fernández FragaThe main aim of this project is to characterise the role of the acetylation of Lys16 of the H4 in neural differentiation. To this end it is planned to study its levels in general and its nuclear distribution, determining total acetylomes in connection with the degree of cellular differentiation, its genetic expression profiles and the degree of chromatin compacting in pluripotent cells and differentiated cells in different neural lines.

From the start of this project work has taken place towards two partial objectives:

  1. The preparation of an in vitro neural differentiation protocol. For this, work commenced using the embryonic mouse cell line JM8A3, and a protocol was followed that basically consists of 3 parts: the growth of the cells in a matrix of embryonic mouse fibroblasts; the subsequent formation of neural precursors by means of culture in suspension with retinoic acid and, finally, differentiation into neurons in a matrix of poly-D-ornithine and laminine. In each of these points, overall acetylation in H4K16ac was analysed by means of HPCE, the results of which showed higher levels of acetylation in lysine 16 of histone H4 in the most differentiated cells.
  2. The preparation of a ChiP-seq protocol to analyse the distribution of the chromatin of H4K16ac. ChIP-Seq is a highly interesting tool for the study of interactions between DNA and proteins, although it is a complex assay. For this reason, it is planned to draw up a protocol that optimises the results of ChIP-Seq in the study of H4K16ac within the experimental system. To identify the conditions of ChIP which make it possible to obtain optimum DNA fragments for use in ultrasequencing, a comparative study was made of several of the key points of the technique: the number of cells at the start, the intercrossing of DNA and histones, chromatin fragmentation, the method of DNA extraction and the antibody used in immunoprecipitation. Following preparation, analysis is now taking place of the acetylation pattern of H4K16 in pluripotent and differentiated cells. There are now fragments of immunoprecipitated DNA that will be ultrasequenced in forthcoming weeks on a Hi-Seq 2000 Illumina platform.

Scientific production
5 articles published in Journals
2 papers at national conferences
7 papers at international conferences

Mario Fernández Fraga

Mario Fernández Fraga is a Tenured Scientific Member of the Superior Science Research Council (CSIC) of the National Biotechnology Centre (CNB), Madrid, and is Head of the Cancer Epigenetics Unit at the Principality of Asturias University Cancer Institute (IUOPA) in the Asturias Central University Hospital (HUCA). Mario F. Fraga was born in Asturias in 1971 and graduated in Molecular Biology from the Complutense University, Madrid, in 1994 and in Biochemistry from the University of Oviedo in 1996. In 2000, he obtained his Doctorate cum laude in the University of Oviedo with a work on developmental epigenetics. In 2001, he started his postdoctoral work in the Cancer Epigenetics Laboratory of the Molecular Pathology Programme of the National Cancer Research Centre (CNIO), as a Fulbright Scholar, and in 2005 he became a Ramón y Cajal researcher in the same group. In 2006, he established a line of research into epigenetics, development, ageing and cancer. Since the beginning of his work as an independent researcher, Mario Fernández Fraga has received uninterrupted financing from the Health Research Fund (Fondo de Investigaciones Sanitarias, FIS) as well as other national and international financing agencies.

Mario Fernández Fraga is the author of more than 120 works published in international journals covering molecular genetics, medicine, biochemistry and cancer. He has written more than 20 chapters of books on epigenetics, development and cancer. He is an editor and reviewer of several international scientific publications, a member of several scientific associations, a habitual reviewer for national and international research evaluation agencies, and is also a member of Spanish and European scientific committees.

*All intellectual property rights belong to the author. Reproduction of all or part of the work without permission from the author is prohibited.
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