PEPTIDE NUCLEIC ACIDS (PNA)-MEDIATED GENE EXPRESSION

描述：（来自申请人的摘要）许多疾病是由 突变导致有缺陷的基因产物或干扰 导致不适当合成的基因表达调节。人类 细胞含有许多结构或功能相关的基因，可以补偿细胞内的蛋白质。 突变基因产物的功能。不幸的是，这些基因中的大多数 在特定组织中表达，或受发育调控， 上游组分的功能可能由于 基因突变。 这个项目的最终目标是诱导这些表达 功能相关基因，并将其应用于人类疾病的基因治疗， 新技术，肽核酸（PNA）诱导的基因表达。PNAS 是具有修饰骨架的合成寡核苷酸。PNA可以结合到 在PNA结合位点的双链结构。这种D环结构具有 被发现具有启动基因转录的能力。在PI的 先前的研究表明，PNA与γ-珠蛋白基因中的靶位点的结合是 发现诱导GFP报告基因和内源性γ-珠蛋白基因 表情然而，PNA结合诱导基因的分子机制 表达没有被研究过。影响PNA介导的细胞凋亡的参数 也没有研究体内基因表达。 在这项提案中，主要研究者计划研究分子 PNA结合诱导基因表达的机制。D环长度 将定义基因转录起始的要求。的 参与PNA结合诱导的基因表达的转录组分将 被确定。来自PNA启动子的RNA转录物的结构将 被研究。PNA结合介导的基因表达与 将研究DNA修复/复制。体内影响参数 PNA基因诱导，例如PNA递送效率和细胞内PNA 还将研究约束力。从这些研究中获得的知识将 有助于了解PNA结合诱导基因的分子基础 表情从这些研究中获得的知识也将用于 探讨利用PNA技术诱导两种基因表达的可能性， 用于治疗两种最常见的人类疾病的人类基因： 镰状细胞病的γ-珠蛋白基因表达和p21的诱导 癌症的基因表达。从该项目中获得的知识也将 有助于理解PNA结合诱导基因表达的分子机制 并为许多人类疾病的治疗提供了直接证据， 遗传疾病和癌症。其目标是： 探讨PNA诱导基因表达的分子机制。 为了研究影响PNA递送效率和细胞内 PNA结合。 探讨PNA结合诱导内源性人基因的可能性 表情

DESCRIPTION: (from the applicant's abstract) Many diseases are caused by mutations that lead to defective gene products or by disturbance in the regulation of gene expression that results in inappropriate synthesis. Human cells contain many structure- or function-related genes that can compensate the function of the mutant gene product. Unfortunately, most of these genes are either developmentally regulated, expressed in specific tissues, or regulated in a pathway by an upstream component whose function may be lost due to mutations in the gene. The ultimate goal of this project is to induce the expression of these function-related genes and apply it to gene therapy of human diseases using a novel technology, peptide nucleic acids (PNAs)-induced gene expression. PNAs are synthetic oligonucleotides with modified backbones. PNAs can bind to double-stranded structures at the PNA binding site. This D-loop structure has been found to have the capability to initiate gene transcription. In the PI's previous studies, binding of PNAs to a target site in the gamma-globin gene was found to induce both the GFP reporter gene and endogenous gamma-globin gene expression. However, the molecular mechanisms of PNA binding-induced gene expression have not been studied. The parameters that affect PNA-mediated in vivo gene expression also have not been investigated. In this proposal the principal investigator plans to study the molecular mechanisms of PNA binding-induced gene expression. The D-loop length requirement for gene transcription initiation will be defined. The transcription components involved in PNA binding-induced gene expression will be determined. The structure of the RNA transcripts from the PNA promoter will be studied. The correlation between PNA binding-mediated gene expression and DNA repair/replication will be investigated. The parameters affecting in vivo PNA gene induction such as PNA delivery efficiency and intracellular PNA binding also will be studied. The knowledge obtained from these studies will contribute to understanding the molecular basis of PNA binding-induced gene expression. The knowledge obtained from these studies will also be used to explore the possibility of using PNA technology to induce the expression of two human genes for treatment of two of the most common human diseases: induction of gamma-globin gene expression for sickle cell disease and induction of p21 gene expression for cancer. The knowledge obtained from this project will also help understand the molecular mechanisms of PNA binding-induced gene expression and provide direct evidence for treatment of many human diseases including genetic diseases and cancer. The aims are: To determine the molecular mechanisms of PNA-induced gene expression. To investigate parameters that affect PNA delivery efficiency and intracellular PNA binding. To explore the possibility of PNA binding-induced endogenous human gene expression.