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.

该提案的目的是开发和验证一个新颖的平台，以丰富完整的曲目 圆形RNA（CIRCRNA）。自从发现> 40年前发现以来，已经有成千上万的circrnas报道 来自人类转录组。可以从它们广泛的事实中衡量circrnas的重要性 在真核细胞中表现出细胞和组织型特异性。他们的表情通常是保守的 跨物种。 Circrnas与神经系统疾病，先天免疫，肝细胞相关 癌和肺纤维化表明它们在细胞生理和病理途径中的作用。这 据报道，目前的方法在富集circrnas曲目方面存在偏见。鉴于它们在各种重要性 疾病，对有效的Circrna隔离技术存在紧急且未满足的需求。在I阶段应用中， 我们提出了两个特定的目的，以开发快速circrna平台，验证并将其功效与当前的功效进行比较 方法。在AIM-1中，我们将在各种条件和细胞类型中开发和建立快速核心平台 获得均匀的circrnas的均匀富集。在AIM-2中，我们将比较快速circrna的功效 在各种细胞条件下采用已知方法。我们认为，I期建议将提供基准 对于II期，它可以简化具有高可重复性的快速ciRCRNA平台 和效率。最后，我们将开发一个快速，坚固且无偏的平台，用于循环RNA隔离， 发现和分析。