Base-resolution sequencing of 6mA in eukaryotic DNA

真核 DNA 中 6mA 的碱基分辨率测序

• 批准号: 9769836
• 负责人: Eric Lieberman Greer
• 金额: $ 22.13万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9769836
• 关键词: Adenine; Antibodies; Bacteria; Bacterial DNA; Base Pairing; Binding; Biological Models; Biological Process; Biology; Buffers; Chromatin Structure; Complement; Coupled; Cytosine; DNA; DNA Methylation; DNA Modification Process; DNA Sequence; Deaminase; Deamination; Deoxyadenosines; Development; Directed Molecular Evolution; Enzyme Tests; Enzymes; Epigenetic Process; Escherichia coli; Eukaryota; Event; Gene Expression; Generations; Genome; Genomic DNA; Genomics; Guanine; High Pressure Liquid Chromatography; Immunoprecipitation; Inosine; Kinetics; Location; Maps; Mass Spectrum Analysis; Measures; Methods; Methylation; Methyltransferase; Modification; Mutagenesis; Mutation; Nucleotides; Organism; Play; Process; Regulation; Research Personnel; Resolution; Role; Scientist; Site; Specificity; Techniques; Testing; Transcription Initiation Site; Uracil; Zebrafish; base; bisulfite sequencing; cost; epigenomics; experimental study; genome wide methylation; imprint; mutant; non-genetic; novel; single molecule real time sequencing; tool; transmission process; zebrafish development

N6-methyladenine (6mA) was recently identified as a new base modification in metazoan genomic DNA. Methylation on adenine was previously thought to be restricted to unicellular organisms. The identification of this mark in metazoans has raised questions about the role of 6mA in metazoan biology. Some studies suggest that 6mA may correlate with developmental regulation, active transcription start sites, or transposon expression, and potentially carry non-genetic information across generations. However, 6mA is a rare DNA modification in eukaryotes and its role in regulating biological processes is still unclear. Because of the low level of 6mA in eukaryotes, some researchers have questioned whether it plays a role in regulating biological processes or is an evolutionary holdover from bacteria. Studies of 6mA have been slowed by a lack of practical methods for accurately examining 6mA locations. Current epigenomic mapping techniques are either prohibitively expensive or do not provide sufficient resolution and accuracy to map the rare 6mA occurrences in metazoan genomes. Here we propose to develop a new technique for identifying sites of adenine methylation genome-wide at nucleotide resolution in a cost-effect manner. We will determine the utility of this technique first in bacteria and then apply it to examine 6mA distribution during development in zebrafish, where we have found that it changes dramatically. This technique will provide a new tool for scientists to define 6mA sites in metazoans to enable the study of its potential role in epigenetics, imprinting and development.

N6-甲基趋化（6MA）最近被确定为后生基因组中的新基础修饰 脱氧核糖核酸。以前认为腺嘌呤上的甲基化仅限于单细胞生物。这 在后生动物中识别此商标已经提出了有关6MA在后生动物中的作用的疑问 生物学。一些研究表明，6MA可能与发育调节，主动调节相关 转录启动位点或转座子表达，并可能携带非遗传信息 几代人。但是，6MA是真核生物中罕见的DNA修饰及其在调节中的作用 生物过程尚不清楚。由于真核生物的6mA水平低，一些研究人员 已经质疑它在调节生物过程中起作用还是进化 从细菌中保留。缺乏实用方法来减慢6MA的研究 准确检查6MA位置。当前的表观基因组映射技术要么是过时的 昂贵或不提供足够的分辨率和准确性来绘制罕见的6mA发生 后生基因组。在这里，我们建议开发一种新技术来识别腺嘌呤的位置 核苷酸分辨率的全基因组以成本效应的方式进行。我们将确定 该技术在细菌中首先的实用性，然后将其应用于检查6MA分布。 斑马鱼的发展，我们发现它发生了巨大变化。这项技术会 为科学家提供了一个新工具，以定义后生动物中的6mA站点，以便研究其潜力 在表观遗传学，印迹和发展中的作用。