Engineering locus-specific binders to DNA modifications

工程化位点特异性结合剂以进行 DNA 修饰

基本信息

批准号：
10593668
负责人：
Albert Keung
金额：
$ 22.8万
依托单位：
NORTH CAROLINA STATE UNIVERSITY RALEIGH
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：
2023-02-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10593668
关键词：
Address Affinity Automobile Driving Awareness BCAR1 gene Base Pairing Binding Binding Proteins Biochemical Biological Biological Assay Biotechnology Cell physiology Cells Chemicals Clustered Regularly Interspaced Short Palindromic Repeats DNA DNA Binding DNA Modification Process DNA Restriction-Modification Enzymes Detection Development Diagnostic Dimensions Directed Molecular Evolution Discipline Disease Engineering Epigenetic Process Etiology Eukaryota Event Exhibits Gene Expression Gene Expression Regulation Genes Genetic Genetic Engineering Genome Genomic DNA Genomics Hour Immunologics Immunoprecipitation Life Location Major Groove Maps Measurement Measures Methylation Modification Molecular Mutation Nature Nucleic Acids Nucleotides Pattern Phenotype Prokaryotic Cells Protein Engineering Proteins RNA Splicing Reagent Reporting Role Safety Sensitivity and Specificity Site Specificity Stimulus Structure Surface System Technology Therapeutic Tissues Work Yeasts Zinc Fingers biological research biological systems bisulfite sequencing cell type epigenome epigenome editing experimental study genome editing genomic locus imprint innovation insight nanopore novel tool whole genome yeast two hybrid system

项目摘要

Project Summary Chemical modifications to genomic DNA are ubiquitous across all kingdoms of life. They are diverse in nature including methylation, hydroxymethylation, and formylation of nucleic acid residues. It is also increasingly clear that these modifications have diverse regulatory roles. In eukaryotes, DNA modifications regulate gene expression, RNA splicing, genome organization, and gene imprinting; these molecular functions also have important downstream consequences for development, disease, and organismal interactions with environmental stimuli and chemicals. Furthermore, with the advent of genome and epigenome editing technologies, these modifications can bias the specificity and efficiency of molecular tools. Given the broad importance of DNA modifications, any experimental tools that are able to control, measure, sense, or track them would have profound impacts across biological disciplines. What is particularly needed, but not yet technologically possibly, is the ability to simultaneously sense epigenetic modifications at specific genomic locations in living and single cells. This capability would unlock a broad palette of new experimental approaches to reveal new insights into topics such as gene imprinting, causation vs. correlation of epigenetic modifications, and cell and tissue stochasticity. Furthermore, it would inform the development of new classes of affinity reagents for biochemical assays, disease detection, and diagnostics and of genome and epigenome editing technologies that are aware of and specific towards DNA modification state. To drive this new realm of studies, we will create the first molecular tool that binds DNA hydroxymethylation at specific and programmable genomic loci, and provide a broadly applicable platform for engineering many other similar molecular reagents.

项目摘要在生活的所有王国中，对基因组DNA的化学修饰都是普遍存在的。他们本质上是多样的包括甲基化，羟甲基和核酸残基的甲基化。也越来越清楚这些修改具有不同的监管角色。在真核生物中，DNA修饰调节基因表达， RNA剪接，基因组组织和基因印记；这些分子功能也具有重要的与环境刺激的发展，疾病和生物相互作用的下游后果化学物质。此外，随着基因组和表观基因组编辑技术的出现，这些修饰可以偏向分子工具的特异性和效率。考虑到DNA修饰的广泛重要性，任何能够控制，衡量，感知或跟踪它们的实验工具将产生深远的影响生物学科。特别需要的，但在技术上尚不具备的是能力同时在生物和单个细胞中特定的基因组位置进行表观遗传修饰。这功能将解锁广泛的新实验方法的调色板，以揭示对诸如等主题的新见解基因印迹，因果关系与表观遗传修饰的相关性以及细胞和组织随机性。此外，它将为生化测定，疾病的新型亲和力试剂的开发提供信息发现和特定的检测，诊断和基因组和表观基因组编辑技术朝向DNA修饰状态。为了推动这一新的研究领域，我们将创建第一个分子工具在特定和可编程基因组基因座上结合DNA羟甲基化，并提供广泛适用的工程许多其他类似的分子试剂的平台。