Sequencing the Mono-Methylated Derivatives of Cytidine
胞苷单甲基化衍生物的测序
基本信息
- 批准号:10581093
- 负责人:
- 金额:$ 40万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2023
- 资助国家:美国
- 起止时间:2023-02-13 至 2025-01-31
- 项目状态:未结题
- 来源:
- 关键词:BenchmarkingBiologicalBiological MarkersCell LineCellsChargeChemicalsClinicalCommunitiesComplexCustomCyclodextrinsCytidineCytosineDefectDerivation procedureDetectionDevelopmentDevicesDiagnosticDiameterDimensionsDiseaseEczemaElectrodesElectrostaticsEnsureEvaluationExonucleaseExoribonucleasesExposure toGoalsHeart failureHeat-Shock ResponseHemolysinHumanInflammatoryIntelligenceIsomerismLengthLipid BilayersMalignant NeoplasmsMalignant neoplasm of cervix uteriMalignant neoplasm of liverMapsMeasurementMetabolic DiseasesMethodologyMethodsMethylationMethyltransferaseMicronutrientsModificationMolecularNational Human Genome Research InstituteNucleotidesPhaseRNAReaderResearchRibonucleotidesRibosomal RNASamplingSmall Business Innovation Research GrantStressSurfaceTargeted ResearchTechnologyTherapeuticTherapeutic InterventionTimeTranslational ResearchVirus Diseasesaccess restrictionsbasecancer celldirected differentiationepitranscriptomicshigh rewardhigh riskinnovationmutantnanonanoporeneurodevelopmentnovelnovel diagnosticspersonalized medicineprecision medicinepreventprognosticprototypesequencing platformtranscriptome sequencingvoltage
项目摘要
Project Summary
The goal of this project is to develop a prototype sequencing platform and the associated methodology capable
of directly sequencing the mono-methylated derivatives of cytosine. Recently, LC-MS/MS methods to assess the
presence of the mono-methylated isomers of cytosine, i.e., 5-methylcytosine (m5C), 4-methylcytidine (m4C), 3-
methylcytidine (m3C), 2`-O-methylcytidine (Cm), as well as the bismethyl derivative 5-methyl-2`-O-methylcytidine
(m5Cm), have made promising discoveries that these modifications change when cells are exposed to oxidative
and inflammatory stress, micronutrient stress, and heat shock stress. Moreover, the placement and extent of
methylated cytosine RNA nucleotides is now believed to be important in neurodevelopment, appears to impact
intelligence, and changes with cancer, eczema, and metabolic disorders. However, no technology currently
exists that is capable of sequencing and quantifying these chemical marks, severely limiting the field’s
understanding and potential utilization of these modifications as biomarkers (for diagnostics, prognostics, and
therapeutic purposes, as examples). In order to enable the proposed feat, during this project, we will specifically
be developing a novel nanopore-based reader with the sensitivity to directly differentiate the mono-methylated
isomers of cytosine as well as the associated sequencing platform for semi-automated, stable, high-accuracy
sequencing measurements. We will then demonstrate and optimize the developed sequencing methodology
through various sequencing demonstrations, including sequencing and mapping the mono-methylated derivates
of cytosine across RNA from two different cell lines. Upon the completion of this project, we will have developed,
optimized, and fully demonstrated the world’s first functioning prototype sequencer capable of directly
sequencing the mono-methylated derivatives of cytosine.
项目摘要
该项目的目的是开发一个原型测序平台和相关的方法论
直接测序胞嘧啶的单甲基化衍生物。最近,LC-MS/MS方法评估
胞嘧啶的单甲基化异构体的存在,即5-甲基胞嘧啶(M5C),4-甲基胞苷(M4C),3--
甲基胞苷(M3C),2`-O-甲基胞丁胺(CM)以及二甲基衍生物5-甲基-2`-O-o-甲基胞丁胺
(M5cm)已承诺发现,当细胞暴露于氧化时,这些修饰发生了变化
以及炎症应激,微量营养素应力和热休克应激。此外,位置和范围
甲基化的胞嘧啶RNA核苷酸现在被认为在神经发育中很重要,似乎会影响
智力以及癌症,湿疹和代谢障碍的变化。但是,目前没有技术
存在能够测序和量化这些化学标记的能力,严重限制了该领域的
理解和潜在利用这些修饰为生物标志物(用于诊断,预后和
治疗目的,例如)。为了实现拟议的壮举,在这个项目中,我们将特别
正在开发一种新型的基于纳米孔的读取器,其灵敏度直接区分单甲基化
胞嘧啶的异构体以及半自动化,稳定,高精度的相关测序平台
测序测量。然后,我们将演示并优化开发的测序方法
通过各种测序演示,包括测序和映射单甲基化衍生物
来自两个不同细胞系的RNA的胞嘧啶。该项目完成后,我们将开发,
优化并充分展示了世界上第一个功能序列的音序器,能够直接
测序胞嘧啶的单甲基化衍生物。
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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Eric Ervin其他文献
Eric Ervin的其他文献
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