Development of Cell-Permeable Peptide Nucleic Acid

细胞渗透性肽核酸的开发

• 批准号: 7408081
• 负责人: DANITH H LY
• 金额: $ 20.49万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7408081
• 关键词: Address; Animal Model; Animals; Basic Science; Binding; Biological; Biomedical Research; Cell Line; Cell Proliferation; Cells; Class; Cues; DNA; Development; Diagnostic; Embryo; Future; Gene Expression; Genes; Genetic Transcription; Hereditary Disease; Human; Human Development; Knowledge; Longevity; Malignant Neoplasms; Mammalian Cell; Methods; Molecular; Nucleic Acids; Oligonucleotides; Peptide Nucleic Acids; Property; RNA; Range; Regenerative Medicine; Research; Site; Stem cells; Technology; Therapeutic Agents; Toxic effect; Transcript; Translations; cancer therapy; cell type; cytotoxicity; design; embryonic stem cell; fascinate; glutarylphenylalanine-4-nitroanilide; human embryonic stem cell; in utero; nucleic acid analog; primitive cell; research study; theories; tool; uptake

DESCRIPTION (provided by applicant): Recently we have developed a new class of peptide nucleic acid called GPNA that is readily taken up by both human somatic and embryonic stem cells, and binds sequence specifically to DNA and RNA. GPNA designed to bind to the transcription and translation start site of the gene transcript elicited potent antisense effect with unusually low cytotoxicity compared to the unmodified PNA and its other derivatives. The proposed research will explore the scope of this particular class of molecules and establish a basic understanding of the factors and mechanisms that control cellular uptake, hybridization and cytotoxicity over a wide range of cell lines, including human ES cells. Specifically, we aim to accomplish four specific objectives within the next five years. Aim 1 (Section D.1): Evaluate the antisense effects of GPNA with human somatic and ES cells; Aim 2 (Section D.2): Optimize cellular uptake, hybridization and cytotoxicity. Aim 3 (Section D.3): Determine the mechanism of GPNA uptake and cytotoxicity. And Aim 4 (Section D.4): Determine the scope of GPNA in regulating gene expression. The proposed study is crucial to the future design and development of nucleic acid mimics for a safe and effective use in animals and in humans. GPNA is, to the best of our knowledge, the first example of nucleic acid analogue taken up by human ES cells. This is intriguing because these primitive cells are extremely difficult to penetrate and are sensitive to the environmental cues. Human ES cells hold the key to understanding early human development that can neither be studied directly in utero nor fully understood with animals model, not to mention its enormous potential for regenerative medicine. Many fascinating questions concerning human ES cell proliferation, differentiation, cellular lifespan and so forth have not yet been addressed. Furthermore, recent evidence suggests that cancers may arise from stem cells. If this proves to be correct, it will alter the course of cancer treatment. Before we can begin to address these questions, an effective method must be developed to regulate gene expression in these and related cell types - this is the aim of our research.

描述（由申请人提供）：最近我们开发了一种新的肽核酸，称为GPNA，它很容易被人类体细胞和胚胎干细胞吸收，并与DNA和RNA特异性结合序列。与未修饰的 PNA 及其其他衍生物相比，设计用于与基因转录物的转录和翻译起始位点结合的 GPNA 引发了有效的反义效应，且细胞毒性异常低。拟议的研究将探索此类特定分子的范围，并建立对控制多种细胞系（包括人类 ES 细胞）的细胞摄取、杂交和细胞毒性的因素和机制的基本了解。具体来说，我们的目标是在未来五年内实现四个具体目标。目标 1（D.1 节）：评估 GPNA 与人体细胞和 ES 细胞的反义效应；目标 2（D.2 节）：优化细胞摄取、杂交和细胞毒性。目标 3（D.3 节）：确定 GPNA 摄取和细胞毒性的机制。目标 4（D.4 节）：确定 GPNA 调节基因表达的范围。 这项研究对于核酸模拟物的未来设计和开发至关重要，以便在动物和人类中安全有效地使用。据我们所知，GPNA 是人类 ES 细胞摄取核酸类似物的第一个例子。这很有趣，因为这些原始细胞极难穿透，而且对环境信号很敏感。人类胚胎干细胞是了解人类早期发育的关键，但它既不能在子宫内直接研究，也不能通过动物模型完全理解，更不用说其在再生医学方面的巨大潜力。关于人类胚胎干细胞增殖、分化、细胞寿命等许多令人着迷的问题尚未得到解决。此外，最近的证据表明癌症可能源自干细胞。如果这被证明是正确的，它将改变癌症治疗的过程。在我们开始解决这些问题之前，必须开发一种有效的方法来调节这些及相关细胞类型中的基因表达——这是我们研究的目的。