GENE THERAPY VECTOR DEVELOPMENT FOR CYSTIC FIBROSIS AND OTHER GENETIC DISEASES

囊性纤维化和其他遗传性疾病的基因治疗载体开发

• 批准号: 2345059
• 负责人: M ROSENFELD
• 金额: --
• 依托单位: NATIONAL HUMAN GENOME RESEARCH INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2345059
• 关键词: DNA replication; DNA replication origin; artificial chromosomes; beta galactosidase; biotechnology; centromere; chloride channels; cystic fibrosis; gene expression; gene therapy; genetic disorder; genetic manipulation; genetic techniques; nucleic acid sequence; reporter genes; telomere; transfection /expression vector

The research goal of the Vector Development Section is to design vectors which overcome problems inherent to current gene transfer approaches such as transient expression (e.g. lipid-mediated plasmid DNA transfer, adenovirus vectors) and the risk for random integration and potential mutagenesis (e.g. retrovirus, adeno-associated virus). An optimal approach would be a vector that provides: (1) stable gene transfer without integration into the host genome, and (2) unlimited capacity to carry DNA so that full length genes can be transferred and analyzed functionally. Toward this end, we are attempting to construct self replicating-vectors in the form of human artificial chromosomes (HAC) which contain human DNA sequences likely necessary for proper chromosome function including telomeres, centromeres and replication origins (Project 1). In addition to the construction of these large vectors, we are also attempting to optimize the delivery of high molecular weight DNA to mammalian cells (Project 2). The primary focus for this study is the cystic fibrosis transmembrane conductance regulator (CFTR) gene contained within in a yeast artificial chromosome (YAC) which also expresses a marker gene Ecoli Lac Z gene [coding for beta-galactosidase]. We are now applying the strategy developed for the CFTR YAC to other YACs and other genetic diseases (Project 3).

向量开发部分的研究目标是设计向量 哪些克服了当前基因转移固有的问题 作为瞬态表达（例如脂质介导的质粒DNA转移， 腺病毒载体）和随机整合和潜力的风险 诱变（例如逆转录病毒，腺相关病毒）。最佳 方法将是提供：（1）稳定基因转移的向量 没有集成到宿主基因组中，（2）无限能力 携带DNA，以便可以转移和分析全长基因 在功能上。为此，我们试图建立自我 人造染色体形式的复制向量（HAC） 其中包含人类DNA序列可能需要适当的染色体 功能包括端粒，中心和复制起源 （项目1）。除了构造这些大量向量外，我们还 还试图优化高分子量DNA的递送 到哺乳动物细胞（项目2）。这项研究的主要重点是 囊性纤维化跨膜电导调节剂（CFTR）基因包含 在酵母人造染色体（YAC）中，也表达 标记基因ecoli lac z基因[编码β-半乳糖苷酶]。我们现在 将为CFTR YAC制定的策略应用于其他YAC和其他 遗传疾病（项目3）。