| 项目编号: | 1505793
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| 项目名称: | UNS: Collaborative Research: Targeted CpG Methylation |
| 作者: | Carl Novina
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| 承担单位: | Dana-Farber Cancer Institute
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| 批准年: | 2014
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| 开始日期: | 2015-06-15
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| 结束日期: | 2018-05-31
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| 资助金额: | USD299997
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| 资助来源: | US-NSF
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| 项目类别: | Standard Grant
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| 国家: | US
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| 语种: | 英语
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| 特色学科分类: | Engineering - Chemical, Bioengineering, Environmental, and Transport Systems
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| 英文关键词: | cpg methylation
; research
; other researcher
; target cpg site
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| 英文摘要: | 1505793/1510652
Ostermeier/Novina
Every cell in the body contains the same DNA sequence. However, the different types of cells in the body (brain, liver, blood, skin, etc.) perform different functions. For example, certain genes that are 'off' in one cell type might be 'on' in another, which enables each cell's unique function. Whether a gene is off or on in a particular cell is due in part to differences in a modification of DNA called CpG methylation. Additionally, certain diseases are caused by abnormal CpG methylation. In this research, a protein will be developed that can alter CpG methylation in a very precise and controlled fashion. This protein will provide other researchers with a tool to study CpG methylation and its effects, to alter cell behavior, and to develop therapies based on the selective altering of a single gene function. A pre-doctoral student and a postdoctoral fellow will be trained in research while performing this research.
Modular, targeted cytosine DNA methyltransferases (MTases) will be developed that are capable of methylating a target CpG site (i.e. 5'-CG-3') at >95% efficiency while leaving non-target CpG sites unmethylated (< 1%). High specificity will be achieved by requiring that the MTase assembles into a functional form at the desired target site in a sequence-dependent manner. These MTases will be optimized using directed evolution. An E. coli reverse selection system will be developed for the rapid assessment of methylation specificity. To adapt the targeted MTases for the repression of mammalian promoters, a user-friendly platform for rapidly identifying CpGs whose methylation leads to repression of eukaryotic promoters will be developed. This work will result in a pipeline of assays and reagents for identifying repressive promoter CpG methylation mediated silencing of endogenous promoters in eukaryotes. The pipeline will be optimized for maximal on-target and minimal off-target gene repression and will be scalable for genome-wide applications. Our research activities will empower researchers to: (1) study methylation's effect on transcription, (2) investigate the mechanisms of spreading of methylation patterns and epigenetic regulation, (3) use targeted MTases as a tool for silencing genes of interest or controlled differentiation of stem cells, and (4) develop targeted MTases as therapeutic agents for the selective silencing of genes (e.g. cancers or viral infections).
This award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the Cellular Dynamics and Function Program of the Division of Molecular and Cellular Biology. |
| 资源类型: | 项目
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| 标识符: | http://119.78.100.158/handle/2HF3EXSE/94387
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| Appears in Collections: | 影响、适应和脆弱性
气候减缓与适应
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| Recommended Citation: | |
Carl Novina. UNS: Collaborative Research: Targeted CpG Methylation. 2014-01-01.
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