SITE SPECIFIC GENE INSERTION BY TRANSPOSITION

通过转座进行位点特异性基因插入

• 批准号: 6342058
• 负责人: ERIC WICKSTROM
• 金额: $ 24.07万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2003-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6342058
• 关键词: bacterial proteins; cell free system; chromatography; chromosomes; electroporation; eukaryote; fungal genetics; gene expression; gene therapy; genes; genetically modified animals; genome; laboratory mouse; microinjections; nucleic acid sequence; plasmids; protein binding; reporter genes; site directed mutagenesis; tissue /cell culture; transfection /expression vector; transposon /insertion element; yeasts; bacterial proteins; cell free system; chromatography; chromosomes; electroporation; eukaryote; fungal genetics; gene expression; gene therapy; genes; genetically modified animals; genome; laboratory mouse; microinjections; nucleic acid sequence; plasmids; protein binding; reporter genes; site directed mutagenesis; tissue /cell culture; transfection /expression vector; transposon /insertion element; yeasts

DESCRIPTION (Applicant's Abstract): The central hypothesis of this proposal is that site and sequence specific insertion of genes into eukaryotic chromosomes can be carried out with Tn7 transpositional proteins. The overall objective is to establish a biological basis for gene therapy based on the use of transposon Tn7 proteins. Tn7 was selected for its ability to integrate into a DNA sequence with specificity and efficiency. Preliminary data support a hypothesis that Tn7 based insertion could provide a safe and efficient method to incorporate therapeutic genes into chromosomes. The central hypothesis will be tested in three Specific Aims: 1.Will Tn7 proteins transposed into eukaryotic targets in vitro? 2. Will Tn7 proteins transpose into eukaryotic targets in vivo? 3. Will Tn7 transposition disrupt normal glutamine-fructose-6-phosphate transaminase (GFPT) expression?

描述（申请人的摘要）： 该提议的中心假设是该地点和序列 可以将基因插入真核染色体中 与TN7转置蛋白一起使用。总体目标是 基于使用 转座子TN7蛋白。 TN7的整合能力被选中 具有特异性和效率的DNA序列。初步数据 支持一个假设，即基于TN7的插入可以提供安全和 将治疗基因纳入染色体的有效方法。这 中央假设将以三个特定目的进行检验：1.将会TN7 蛋白质在体外转移到真核靶标？ 2。Will TN7 蛋白质在体内转移到真核靶标？ 3。Will TN7 转座破坏正常谷氨酰胺 - 果糖-6-磷酸转氨酸酶 （GFPT）表达？