Over the past decade, epigenetic phenomena claimed a central role in cell regulatory processes and proved important factors for understanding complex human diseases. One of the best understood epigenetic mechanisms is DNA methylation. In the mammalian genome, cytosines (C) were long known to exist in two functional states: unmethylated or methylated at the 5-position of the pyrimidine ring (5mC). Recent studies of genomic DNA from the human and mouse brain, neurons and from mouse embryonic stem cells found that a substantial fraction of 5mC in CpG dinucleotides is converted to 5-hydroxymethylcytosine (hmC) by the action of oxygenases. These findings provided important clues in a long elusive mechanism of active DNA demethylation and bolstered a fresh wave of studies in the area of epigenetic regulation in mammals. However, such studies are hindered by the shortcomings of available experimental techniques for genome-wide epigenome analysis. This project is dedicated to developing a break-through technique for genome-wide analysis of the modification status of CpG dinucleotides that combines single-base resolution with targeted and economic sequencing of the genome.

Public Health Relevance

Changes in cytosine modification patterns in the genome may predispose individuals to various human diseases, including cancer, schizophrenia, autism, asthma, and diabetes. Research into the epigenetic misregulation in human disease is hampered by the lack of high resolution and cost-effective techniques. Our new technologies for single-nucleotide analysis of cytosine modifications will be an indispensible addition to the tool box of comprehensive epigenomic studies.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21HG007200-02
Application #
8651509
Study Section
Genomics, Computational Biology and Technology Study Section (GCAT)
Program Officer
Smith, Michael
Project Start
2013-04-12
Project End
2015-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Vilniaus Universitetas
Department
Type
DUNS #
City
Vilnius
State
Country
Lithuania
Zip Code
Osipenko, Aleksandr; Plotnikova, Alexandra; Nainyt?, Milda et al. (2017) Oligonucleotide-Addressed Covalent 3'-Terminal Derivatization of Small RNA Strands for Enrichment and Visualization. Angew Chem Int Ed Engl 56:6507-6510
Staševskij, Zdislav; Gibas, Povilas; Gordevi?ius, Juozas et al. (2017) Tethered Oligonucleotide-Primed Sequencing, TOP-Seq: A High-Resolution Economical Approach for DNA Epigenome Profiling. Mol Cell 65:554-564.e6
Tomkuvien?, Migl?; Kriukien?, Edita; Klimašauskas, Saulius (2016) DNA Labeling Using DNA Methyltransferases. Adv Exp Med Biol 945:511-535
Masevi?ius, Viktoras; Nainyt?, Milda; Klimašauskas, Saulius (2016) Synthesis of S-Adenosyl-L-Methionine Analogs with Extended Transferable Groups for Methyltransferase-Directed Labeling of DNA and RNA. Curr Protoc Nucleic Acid Chem 64:1.36.1-13