Real-time PCR expression profiling of microRNA

microRNA 的实时 PCR 表达谱

• 批准号: 7808845
• 负责人: THOMAS D. SCHMITTGEN
• 金额: $ 20.91万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-25 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7808845
• 关键词: 3' Untranslated Regions; Area; Attention; Award; Binding; Biological; Biological Assay; Biological Process; Cancerous; Cells; Clinical; Code; Complementary DNA; Disease; Evaluation; Fibrinogen; Formalin; Freezing; Gene Expression; Gene Expression Regulation; Grant; Health; Homeostasis; Human; Length; Liver; Malignant Neoplasms; Malignant neoplasm of pancreas; Messenger RNA; Methodology; MicroRNAs; Molecular Profiling; Monitor; Normal tissue morphology; Nucleotides; Pancreas; Paraffin Embedding; Proteins; RNA; RNA Binding; Research; Research Personnel; Role; Sampling; Sensitivity and Specificity; Small Interfering RNA; Specificity; Technology; Time; Tissues; carcinogenesis; laser capture microdissection; pre-miRNA; programs

DESCRIPTION (provided by applicant): microRNA is a newly discovered class of endogenous, small interfering RNA. MicroRNA binds to messenger RNA and translationally represses protein levels. While over 300 microRNAs have been discovered in humans alone, their biological function, targets, expression levels and role in disease remain largely unknown. A role between microRNA expression and carcinogenesis has been proposed. There is a lack of sensitive, high-throughput methodologies to monitor the expression of microRNAs. microRNA are challenging molecules to quantify because the microRNA precursors consists as a stable hairpin and the mature microRNA is only 22 nucleotides in length. We propose to evaluate sensitive and specific real-time PCR assays to quantify the expression of the mature and microRNA precursors. The microRNA expression will be analyzed in a number of important biological conditions relating to human cancer. The microRNA expression will be determined in specific sections of cancer and normal tissue isolated by laser-capture microdissection. The expression of mature and precursor microRNAs will be compared to using real-time PCR and a cDNA micro array. microRNA expression will be studied in clinical samples of human pancreatic cancer. The unparallel sensitivity and specificity of real-time PCR as applied to this new and exciting class of regulatory RNAs should propel the field into new directions not only in cancer but in other areas of human health.

描述（由申请人提供）：microRNA是一类新发现的内源性小干扰RNA。 MicroRNA 与信使 RNA 结合并翻译抑制蛋白质水平。虽然仅在人类中就发现了 300 多种 microRNA，但它们的生物学功能、靶标、表达水平和在疾病中的作用仍然很大程度上未知。已经提出了 microRNA 表达与癌发生之间的作用。缺乏灵敏、高通量的方法来监测 microRNA 的表达。 microRNA 是一种具有挑战性的分子，因为 microRNA 前体由稳定的发夹组成，而成熟的 microRNA 长度仅为 22 个核苷酸。我们建议评估灵敏且特异的实时 PCR 检测，以量化成熟和 microRNA 前体的表达。将在与人类癌症相关的许多重要生物学条件下分析 microRNA 的表达。将在通过激光捕获显微切割分离的癌症和正常组织的特定切片中测定 microRNA 表达。成熟和前体 microRNA 的表达将与使用实时 PCR 和 cDNA 微阵列进行比较。将在人类胰腺癌的临床样本中研究 microRNA 表达。实时 PCR 无与伦比的灵敏度和特异性应用于这一令人兴奋的新型调节 RNA 领域，不仅会推动该领域进入癌症领域，而且会推动人类健康其他领域的新方向。