Tools for Single Molecule and Single Cell Epigenomic Analysis
单分子和单细胞表观基因组分析工具
基本信息
- 批准号:8534233
- 负责人:
- 金额:$ 54.18万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2012
- 资助国家:美国
- 起止时间:2012-08-21 至 2015-06-30
- 项目状态:已结题
- 来源:
- 关键词:AddressAgreementAntibodiesBasic ScienceBindingBiological AssayBiological ProcessCell SeparationCellsChemicalsChromatinClinical assessmentsComplexComputer-Assisted Image AnalysisCultured CellsCytosineDNADNA SequenceDNA-Binding ProteinsDataData CollectionData SetDevelopmentDevicesEmbryoEngineeringEpigenetic ProcessFluorescent Antibody TechniqueFluorescent ProbesGenerationsGenesGenetic MaterialsGenomeGoalsHealthHistonesHuman BiologyImageImmunoprecipitationIndividualMammalsMeasuresMedicineMethodsMethylationMicroscopeModificationMonitorPatientsPharmaceutical PreparationsPopulationPreparationProcessPropertyPublishingReportingResearchSamplingSorting - Cell MovementSourceSurfaceTechnologyTestingTherapeuticTimebasebisulfitechromatin immunoprecipitationcostdisease diagnosisdrug developmentepigenomeepigenomicsfluorescence activated cell sorter devicefluorophoregenome-widehistone modificationimprovedinduced pluripotent stem cellinformation gatheringnanofluidicnext generationoperationresearch studyresponsesingle moleculetooltraitvoltage
项目摘要
Epigenetic features in mammals include covalent modifications to histones, and methylation of cytosines.
Their proper placement is fundamental to many biological processes. Assaying features in the epigenome is
important for understanding human biology, diagnosing disease, monitoring responses to epigenome
modifying drugs and facilitating development of new medicines. Such assays will also facilitate emerging
therapeutics based on embryonic and induced pluripotent stem cells, which require modifying the cells to
assume epigenetic states of differentiated cells. State-of-the-art assays for histone modifications use
chromatin immunoprecipitation (ChIP), followed by genome wide sequencing. Methylated DNA can be
identified by immunoprecipitation followed by sequencing, or by bisulfite sequencing. There are two
fundamental limitations with all these approaches. First, they query only one epigenetic feature at a time.
Epigenetic features arise in combinations, and those combinations rather than individual features regulate the
underlying genes. Unless multiple features can be detected and measured simultaneously, it is not possible to
know, with certainty, when combinations coexist on a given gene. Second, assays use populations of cells
and report the average epigenetic states within the population, not the actual distribution of epigenetic states
present on individual DNA molecules comprising the population. Third, ChIP often uses abundant amounts of
chromatin making it impractical to assay multiple epigenetic features in rare or impossible to culture cells. In
this application, we seek to develop a transforming technology, Single Chromatin molecule Analysis in
Nanofluidics (SCAN) that can overcome each of these limitations and revolutionize epigenomic studies. In
SCAN, chromatin molecules are bound to fluorescent probes recognizing distinct epigenetic features, then
driven by voltage through nanofluidic channels where the fluorescent properties of single molecules are
detected. By using multiple probes, each recognizing different features and carrying distinct fluorophores, we
can directly detect their binding to individual molecules, allowing precise enumeration of multiple epigenetic
features simultaneously. Our first-generation devices were operated in an analytical mode, simply counting
features. Our second-generation devices were operated in a preparative mode, allowing us to sort and isolate
molecules carrying defined epigenetic features. In this proposal, we seek to further develop this next
generation epigenomics technology. First, we will modify the analytical device and analyte preparation to
increase sample throughput by two orders of magnitude. Second, we will use the new analytical device to
address selected questions in epigenomics. Third, we will use our preparative device to isolate chromatin with
defined epigenetic features, sequence the DNA and compare our results to data obtained by current ChIP-seq
methods.
哺乳动物的表观遗传特征包括组蛋白的共价修饰和胞嘧啶的甲基化。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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HAROLD G CRAIGHEAD其他文献
HAROLD G CRAIGHEAD的其他文献
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{{ truncateString('HAROLD G CRAIGHEAD', 18)}}的其他基金
Adaptable and scalable electroporation for cellular therapy
用于细胞治疗的适应性和可扩展的电穿孔
- 批准号:
10545845 - 财政年份:2022
- 资助金额:
$ 54.18万 - 项目类别:
Tools for Single Molecule and Single Cell Epigenomic Analysis
单分子和单细胞表观基因组分析工具
- 批准号:
8340779 - 财政年份:2012
- 资助金额:
$ 54.18万 - 项目类别:
Tools for Single Molecule and Single Cell Epigenomic Analysis
单分子和单细胞表观基因组分析工具
- 批准号:
8683212 - 财政年份:2012
- 资助金额:
$ 54.18万 - 项目类别:
In vivo Detection and Imaging of Epigenetic Histone Modifications and Modifying E
表观遗传组蛋白修饰和修饰 E 的体内检测和成像
- 批准号:
8269067 - 财政年份:2010
- 资助金额:
$ 54.18万 - 项目类别:
In vivo Detection and Imaging of Epigenetic Histone Modifications and Modifying E
表观遗传组蛋白修饰和修饰 E 的体内检测和成像
- 批准号:
8144800 - 财政年份:2010
- 资助金额:
$ 54.18万 - 项目类别:
In vivo Detection and Imaging of Epigenetic Histone Modifications and Modifying E
表观遗传组蛋白修饰和修饰 E 的体内检测和成像
- 批准号:
8471090 - 财政年份:2010
- 资助金额:
$ 54.18万 - 项目类别:
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