Sequencing the Mono-Methylated Derivatives of Cytidine

胞苷单甲基化衍生物的测序

• 批准号: 10581093
• 负责人: Eric Ervin
• 金额: $ 40万
• 依托单位: ELECTRONIC BIOSCIENCES, INC.
• 依托单位国家: 美国
• 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: https://reporter.nih.gov/project-details/10581093
• 关键词: Benchmarking; Biological; Biological Markers; Cell Line; Cells; Charge; Chemicals; Clinical; Communities; Complex; Custom; Cyclodextrins; Cytidine; Cytosine; Defect; Derivation procedure; Detection; Development; Devices; Diagnostic; Diameter; Dimensions; Disease; Eczema; Electrodes; Electrostatics; Ensure; Evaluation; Exonuclease; Exoribonucleases; Exposure to; Goals; Heart failure; Heat-Shock Response; Hemolysin; Human; Inflammatory; Intelligence; Isomerism; Length; Lipid Bilayers; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Malignant neoplasm of liver; Maps; Measurement; Metabolic Diseases; Methodology; Methods; Methylation; Methyltransferase; Micronutrients; Modification; Molecular; National Human Genome Research Institute; Nucleotides; Phase; RNA; Reader; Research; Ribonucleotides; Ribosomal RNA; Sampling; Small Business Innovation Research Grant; Stress; Surface; Targeted Research; Technology; Therapeutic; Therapeutic Intervention; Time; Translational Research; Virus Diseases; access restrictions; base; cancer cell; directed differentiation; epitranscriptomics; high reward; high risk; innovation; mutant; nano; nanopore; neurodevelopment; novel; novel diagnostics; personalized medicine; precision medicine; prevent; prognostic; prototype; sequencing platform; transcriptome sequencing; voltage

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.

项目总结