SLEEPING BEAUTY TRANSPOSON FOR GENE THERAPY

用于基因治疗的睡美人转座子

• 批准号: 6442591
• 负责人: PERRY B. HACKETT
• 金额: $ 10.62万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6442591
• 关键词: biotechnology; cell line; clinical research; gene therapy; human subject; laboratory mouse; laboratory rat; liposomes; liver cells; mucopolysaccharidosis type I; tissue /cell culture; transfection /expression vector; transposon /insertion element

Non-viral gene transfer procedures are needed for human gene therapy in order to achieve long-term maintenance and expression of newly introduced genes. The purpose of this project is to evaluate the efficacy of the Sleeping Beauty transposon system as a gene transfer vector for gene therapy. The transposon system is binary, consisting of a transposon containing a transgene and a source of transposase enzyme. The transposase is able to mediated enhanced integration of marker/reporter genes in several mammalian culture cell lines in vitro as well as in fertilized vertebrate. Here, the examination of transposon system will be extended to liver cells, ranging from tissue-cultured hepatoma cells, to primary hepatocytes and livers of whole animals. Liposomes complexed with lactosylated DNA condensing agents will be used to deliver the transposon system. Genes that are involved human genetic disease, and for which the transposon system. The successful demonstration of transposition of new sequences into mammalian cells would constitute a proof-of-principle for using this integrating mechanism in humans for gene therapy that does not employ viral vectors.

人类基因治疗需要非病毒基因转移程序，以实现新引入基因的长期维持和表达。本项目的目的是评估睡美人转座子系统作为基因治疗的基因转移载体的功效。转座子系统是二元的，由含有转基因的转座子和转座酶的来源组成。该转座酶能够介导标记/报告基因在几种哺乳动物体外培养细胞系以及受精脊椎动物中的增强整合。在这里，转座子系统的研究将扩展到肝细胞，从组织培养的肝癌细胞，原代肝细胞和整个动物的肝脏。与乳糖基化DNA缩合剂复合的脂质体将用于递送转座子系统。涉及人类遗传疾病的基因，以及转座子系统。将新序列转座到哺乳动物细胞中的成功证明将构成在人类中使用这种整合机制进行不使用病毒载体的基因治疗的原理证明。