Utilizing TALEN technology to regulate human microRNA's

利用TALEN技术调控人类microRNA

• 批准号: 8568549
• 负责人: Ryan M O'Connell
• 金额: $ 223.5万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8568549
• 关键词: Autoimmunity; Binding Sites; Biological Assay; Biology; Cardiovascular Diseases; Code; DNA-Binding Proteins; Disease; Engineering; Gene Deletion; Genes; Germ Lines; Health; Heart Diseases; Human; Immune response; Inflammatory Response; Link; Malignant Neoplasms; Messenger RNA; MicroRNAs; Mus; Nerve Degeneration; Phenotype; Physiology; Proteins; Regulation; Role; Series; Technology; Time; Tumor Suppressor Proteins; Untranslated Regions; combat; homologous recombination; human disease; improved; in vivo; man; novel strategies; nuclease; public health relevance; transcription factor

DESCRIPTION (provided by applicant): MicroRNAs (miRNAs) have quickly emerged as important regulators of mammalian physiology owing to their precise regulation of critical protein coding genes. Importantly, perturbations in expression or function of specific miRNAs has been linked to a plethora of human pathological conditions including cancer, autoimmunity, cardiovascular disease and neurodegeneration. Although miRNA gene deletion in mice via homologous recombination has led to an improved understanding of how they function during times of health and disease, there remains a fundamental need to manipulate miRNA genes and their target binding sites in the human germ line. This would allow for the study of their biology in man, and may be a strategic means by which miRNAs can be targeted therapeutically to combat human disease. We propose to develop a novel approach to activating, repressing or disrupting human miRNA genes in vivo by engineering TALE proteins that will function as transcription factors or nucleases that specifically target miRNA genes or their binding sites in the 3'UTRs of key target mRNAs. We will focus on targeting human miRNA---155, an "oncomiR" involved in regulating immune responses, and miR---146a, a tumor suppressor miRNA that inhibits inflammatory responses. Following proper targeting and disruption of these miRNAs or their binding sites in the 3' UTRs of relevant target mRNAs, a series of functional assays will be carried out to assess the roles of human miR---155 and miR---146a in regulating immune responses and cancer phenotypes. Beyond these two miRNAs, this approach will be compatible with any human miRNA making it a powerful, specific and versatile technology that has not yet been used to study miRNAs.

描述（由申请人提供）：由于其精确调节关键蛋白质编码基因，microRNA（miRNA）已迅速成为哺乳动物生理学的重要调节剂。重要的是，特定miRNA的表达或功能的扰动与众多人类病理状况有关，包括癌症，自身免疫性，心血管疾病和神经变性。尽管小鼠通过同源重组中的miRNA基因缺失导致对它们在健康和疾病时期的功能有了改善的了解，但仍有基本需要操纵miRNA基因及其靶标结合位点。这将允许在人类中研究其生物学，并且可能是一种战略手段，可以将miRNA靶向治疗以打击人类疾病。我们建议通过工程蛋白在体内激活，抑制或破坏人体miRNA基因的新方法，该蛋白将充当转录因子或核酸酶，或在关键靶标mRNA的3'UTR中专门针对miRNA基因或其结合位点的转录因子或核酸酶。我们将专注于靶向人类miRNA - 155，一种参与调节免疫反应的“ oncomir”，以及miR --- 146a，一种抑制炎症反应的肿瘤抑制miRNA。在相关靶标mRNA的3'UTR中对这些miRNA或其结合位点的适当靶向和破坏后，将进行一系列功能测定，以评估人类miR的作用------ 155和miR ----- 146a在调节免疫反应和癌症表型中的调节中。除了这两个miRNA之外，这种方法将与任何人类miRNA兼容，使其成为一种尚未用于研究miRNA的强大，特定和多才多艺的技术。