Real-time PCR expression profiling of microRNA

microRNA 的实时 PCR 表达谱

• 批准号: 7596042
• 负责人: THOMAS D. SCHMITTGEN
• 金额: $ 20.92万
• 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 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7596042
• 关键词: 3' Untranslated Regions; Area; Attention; Award; Binding; Biological; Biological Assay; Biological Process; Cancerous; Cells; Class; Clinical; Code; Complementary DNA; Condition; 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; Numbers; Pancreas; Paraffin Embedding; Polymerase Chain Reaction; Proteins; RNA; RNA Binding; Range; Research; Research Personnel; Role; Sampling; Sensitivity and Specificity; Small Interfering RNA; Specificity; Technology; Time; Tissues; carcinogenesis; laser capture microdissection; pre-miRNA; programs; size

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，小型干扰RNA。 MicroRNA与Messenger RNA结合并翻译抑制蛋白质水平。尽管仅在人类中就发现了300多个microRNA，但它们的生物学功能，靶标，表达水平和疾病的作用在很大程度上未知。已经提出了microRNA表达和癌变之间的作用。缺乏敏感的高通量方法来监测microRNA的表达。 microRNA是挑战分子来量化的，因为microRNA前体作为稳定的发夹组成，成熟的microRNA的长度仅为22个核苷酸。我们建议评估敏感和特定的实时PCR分析，以量化成熟和microRNA前体的表达。将在许多与人类癌症有关的重要生物学条件下分析microRNA表达。 microRNA表达将在通过激光捕获显微解剖分离的癌症和正常组织的特定切片中确定。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。将在人胰腺癌的临床样本中研究microRNA表达。适用于这一新的令人兴奋的监管RNA类的实时PCR的非平行敏感性和特异性不仅可以使该领域不仅在癌症中，而且在人类健康的其他领域中。