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Dnmt3b activities in mouse development

Dnmt3b 在小鼠发育中的活性

基本信息

批准号：
10419773
负责人：
Rene Opavsky
金额：
$ 30.5万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-01 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10419773
关键词：
Affect Assisted Reproductive Technology Birth Cells ChIP-seq Clinical Complex Conceptions DNA DNA Methylation DNA Modification Methylases DNA Transposable Elements DNMT3B gene DNMT3a Data Defect Development Disease Down-Regulation Embryo Embryonic Development Enhancers Enzymes Epiblast Epigenetic Process Ethics Family Fertilization in Vitro Future Gene Expression Gene Expression Regulation Genes Genetic Transcription Genome Genomic Imprinting Genomic Segment Genomics Health Human Human Cell Line Immunoprecipitation In Vitro Individual Knock-out Knockout Mice Knowledge Large Intestine Carcinoma Lead Longitudinal Studies Malignant Epithelial Cell Measures Mediating Medicine Methylation Methyltransferase Modeling Modification Molecular Molecular Analysis Mus Pathogenesis Phenotype Physiological Physiological Processes Play Pre-implantation Embryo Development Preventive care Procedures Process Proteins Reporting Role Techniques Testing Time Tissue-Specific Gene Expression Tissues Totipotency Transcriptional Regulation X Inactivation demethylation embryonic stem cell epigenome genetic approach genome wide methylation genome-wide genomic locus histone modification implantation imprint in vivo methylation pattern mouse development novel therapeutic intervention pluripotency postnatal development prevent promoter recruit transcriptome sequencing

项目摘要

DNA methylation is an epigenetic modification involved in transcriptional regulation of genes involved in development and differentiation, and its deregulation contributes to human pathogenesis. It is catalyzed by the family of DNA methyltransferases including catalytically active Dnmt1, Dnmt3a, Dnmt3b. DNA methylation plays a major role in preimplantation development in mice. To establish a new epigenome, the mouse zygotic genome undergoes epigenetic reprogramming, including global DNA demethylation at the 8-cell stage. Upon implantation, a wave of de novo methylation in epiblast cells mediated by de novo enzymes Dnmt3a and Dnmt3b results in new methylation patterns maintained by Dnmt1 that form a basis for tissue-specific expression and differentiation. Dnmt3b regulates developmental and imprinted genes, X chromosome inactivation, pericentromeric regions, gene bodies and other genomic regions. Its importance in mouse development was demonstrated by embryonic lethality of Dnmt3b-/- mice. We recently found that Dnmt3bCI/CI mice expressing catalytically inactive Dnmt3bCI protein survived both pre- and postnatal development. Molecular analysis suggested that accessory function - the ability to recruit other Dnmts to proper genomic loci – of Dnmt3b rather than its catalytic activity, is important for methylation and survival. Here we hypothesize that Dnmt3b is a multifaceted protein whose various activities involved in methylation affect pre- and postnatal development and are critical to prevent disease formation in mice. To test this hypothesis, in Aim 1 we analyze global methylation and expression at different stages of development in mice lacking various Dnmt activities to determine the scope of Dnmt3b’s accessory function in Dnmt3a-mediated de novo methylation in vivo as well as regulation of transcription of various genomic features including gene bodies, germline genes and transposons. In Aim 2, will test the ability of Dnmt3b to complex with other Dnmts and contribute to de novo methylation induced by other Dnmts in Dnmt1-/-;Dnmt3a-/-;Dnmt3b-/- triple knockout mouse embryonic stem cells. In addition, we will genetically test the importance of Dnmt3a and Dnmt3l for Dnmt3b’s accessory function and validate our data in a human cell line. In Aim 3, we will perform longitudinal study of Dnmt3b+/+ and Dnmt3bCI/CI mice conceived through the use of in vitro fertilization (IVF) technique to analyze disease development, Dnmt levels, the rate of methylation and gene expression errors, as well as their persistence over time. Collectively, our studies will reveal physiological relevance of Dnmt3b activities in mouse development, uncover basic mechanisms utilizing Dnmt3b functions and their involvement in IVF. Our results could result in changes in Assisted Reproductive Technologies (ART) and affect the focus of preventive care for ART- conceived individuals.

DNA甲基化是一种表观遗传修饰，涉及基因的转录调控