cis-Acting Elements Regulating Developmental Control of Replication Timing

调节复制时间发育控制的顺式作用元件

• 批准号: 8425084
• 负责人: David M Gilbert
• 金额: $ 28.65万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8425084
• 关键词: Address; Boundary Elements; Cations; Cell Differentiation process; Cell Nucleus; Cell division; Cells; Characteristics; Chromatin; Chromosome Structures; Chromosomes; DNA; DNA Sequence; DNA biosynthesis; Development; Disease; Dissection; Eating; Elements; Engineering; Epigenetic Process; Euchromatin; Eukaryotic Cell; Genes; Genetic; Genetic Recombination; Genetic Transcription; Genome; Genomics; Goals; Growth and Development function; Heterochromatin; Human; Human Chromosomes; Integrase; Knowledge; Link; Location; Malignant Neoplasms; Mammals; Maps; Mediating; Mission; Modeling; Molecular; Monitor; Mus; Nuclear; Pathogenesis; Public Health; Regulation; Regulatory Element; Replicon; Role; Shipping; Ships; Site; Specific qualifier value; System; Testing; Time; Tissues; Transcriptional Regulation; Work; abstracting; base; cell type; cis acting element; disability; embryonic stem cell; genome-wide; human disease; innovation; insight; novel; programs; promoter; research study; stem cell differentiation; tool

DESCRIPTION (provided by applicant): PROJECT SUMMARY / ABSTRACT Abnormal temporal control of replication is observed in many diseases but causal linkages are unknown. This gap will remain incomprehensible until the mechanisms regulating replication timing during normal development are understood. The long-term goal is to understand the relationship of replication timing to cellular epigenetic states and disease. The immediate goal is to identify cis-acting DNA/chromatin elements that regulate changes in replication timing during differentiation of mouse embryonic stem cells (ESCs). Mouse ESCs are an ideal experimental system due to the availability of chromosome engineering tools, directed cell differentiation systems, and comprehensive genome-wide maps of replication timing and transcription. These maps have identified the molecular coordinates of programmed changes in replication timing that occur in 400-800kb units termed "replication domains". The central hypothesis is that discrete identifiable chromatin or DNA sequence features dictate the boundaries of replication domains and the developmentally induced changes in their replication time. The rationale for this proposal is that identifying DNA/chromatin elements regulating replication timing is the essential next step in elucidating mechanisms regulating replication timing and its relationship to disease. Aim1 will test the hypothesis that replication domains are fundamental units of chromosome structure and function that can be transferred to an ectopic location. Large pieces of cloned genomic DNA from a developmentally regulated replication domain will be introduced into a region of constitutive replication timing. Repli- cation timing of the insert and flanking DNA will be monitored during differentiation to identify the minimal sequences constituting a unit of regulation. Aim2 will distinguish between models in which specific boundary elements punctuate temporally distinct domains vs. models of boundaries as passively replicated chromatin between actively programmed domains. Nested deletions will be engineered in developmentally controlled replication timing transition regions and the effects of deletions on the regulation of replication timing will be determined. Aim3 will test the hypothesis that transcription within a silent late replicating domain initiates a switch to early replication. Promoter and regulatory elements controlling transcription within a developmentally regulated replication domain will be deleted, replaced with an inducible promoter, and the effects of such manipulations on the regulation of replication timing will be analyzed. Studies described here will identify cis-acting elements regulating the developmental control of replication timing. This contribution is significant because identifying regulatory elements of replication timing control is a pre-requisite to understanding the role of replication timing in chromosome-based diseases. The work proposed here is innovative in that it proposes a novel combination of chromosome engineering and directed embryonic stem cell (ESC) differentiation to address the mechanisms eliciting developmentally programmed changes in replication timing.

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