A quick, and robust, and unbiased platform for circular RNAs isolation, discovery and profiling.

一个快速、强大且公正的循环 RNA 分离、发现和分析平台。

• 批准号: 10761203
• 负责人: Prashant K. Khade
• 金额: $ 35.29万
• 依托单位: RIBO-THERAPEUTICS LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-09 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10761203
• 关键词: Acceleration; Address; Affect; Benchmarking; Biochemical; Biochemistry; Bioinformatics; Biology; Biotinylation; Buffers; California; Cell physiology; Cells; Complex; Computational Biology; DNA; Data; Detection; Development; Digestion; Disease; Environment; Eukaryotic Cell; Exons; Gene Expression; Genomics; Goals; Hour; Human; Hydroxyl Radical; Immunity; Introns; Ligation; Malignant Neoplasms; Maps; Medical; Messenger RNA; Methodology; Methods; MicroRNAs; Modification; Natural Immunity; Nature; Pathologic; Pathway interactions; Phase; Physiological; Poly A; Poly(A) Tail; Positioning Attribute; Primary carcinoma of the liver cells; Principal Investigator; Process; Publications; Pulmonary Fibrosis; RNA; RNA Sequences; RNA Splicing; Regulator Genes; Reporting; Reproducibility; Role; Sampling; Sensitivity and Specificity; Specificity; Streptavidin; Structure; Technology; Tissues; Universities; Virus Diseases; bioinformatics pipeline; cell type; cellular pathology; circular RNA; comparative efficacy; comparison control; efficacy evaluation; improved; insight; magnetic beads; nervous system disorder; novel; nuclease; predictive tools; ribonuclease R; transcriptome; transcriptome sequencing

The goal of this proposal is to develop and validate a novel platform for the enrichment of the complete repertoire of circular RNAs (circRNAs). Thousands of circRNAs have been reported since their discovery >40 years ago from human transcriptome. The importance of circRNAs can be gauged from the fact that they are broadly expressed in eukaryotic cells, show cell and tissue-type specificity; and their expression is often conserved across species. CircRNAs have been implicated in neurological diseases, innate immunity, hepatocellular carcinoma and lung fibrosis demonstrating their role in cellular physiological and pathological pathways. The present approaches are reported to be biased in enriching circRNAs repertoire. Given their importance in various diseases, there is an urgent and unmet need for efficient circRNA isolation technology. In Phase-I application, we proposed two specific aims to develop Quick-circRNA platform, validate and compare its efficacy with current methods. In aim-1 we will develop and establish Quick-circRNA platform in various conditions and cell types in obtaining uniform an unbiased enrichment of circRNAs. In aim-2 we will compare the efficacy of Quick-circRNA with known methods in various cellular conditions. We believe that phase-I proposal will provide a benchmark for phase-II, which can streamline the Quick-circRNA platform for enriching circRNAs with high reproducibility and efficiency. In the end, we will develop a quick, and robust, and unbiased platform for circular RNAs isolation, discovery and profiling.

这项提议的目标是开发和验证一个丰富完整剧目的新平台 环状RNA(CircRNAs)。自从40年前发现CircRNA以来，已有数千种CircRNAs被报道 来自人类转录组。CircRNA的重要性可以从它们广泛存在的事实来衡量 在真核细胞中表达，表现出细胞和组织类型的特异性；并且它们的表达通常是保守的 跨物种。CircRNAs与神经系统疾病、先天免疫、肝细胞 癌症和肺纤维化表明它们在细胞生理和病理途径中的作用。这个 据报道，目前的方法在丰富CircRNA谱系方面存在偏见。鉴于它们在不同领域的重要性 随着人类疾病的发展，对有效的CircRNA分离技术的需求迫切且尚未得到满足。在第一阶段应用中， 我们提出了开发Quick-CircRNA平台的两个具体目标，验证了其有效性，并与现有的方法进行了比较 方法：研究方法。在AIM-1中，我们将在不同条件和细胞类型下开发和建立Quick-CircRNA平台 获得均匀和无偏见的CircRNA的丰富。在AIM-2中，我们将比较快速环状RNA的有效性 在各种细胞条件下使用已知的方法。我们相信，第一阶段提案将提供一个基准 对于第二阶段，可以简化Quick-CircRNA平台，以丰富具有高重复性的CircRNA 和效率。最终，我们将开发一个快速、健壮、无偏倚的环状RNA分离平台， 发现和剖析。