SITE SPECIFIC GENE INSERTION BY TRANSPOSITION

通过转座进行位点特异性基因插入

• 批准号: 2745214
• 负责人: ERIC WICKSTROM
• 金额: $ 23.95万
• 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 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2745214
• 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

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）： 该提案的中心假设是位点和序列 可以将基因特异性插入真核染色体 与 Tn7 转座蛋白一起。总体目标是 建立基因治疗的生物学基础 转座子 Tn7 蛋白。 Tn7 因其整合能力而被选中 具有特异性和效率的 DNA 序列。初步数据 支持基于 Tn7 的插入可以提供安全且可靠的假设 将治疗基因整合到染色体中的有效方法。这 中心假设将在三个具体目标中进行检验： 1.Will Tn7 蛋白质在体外转变成真核靶标？ 2. 威尔 Tn7 蛋白质在体内转变成真核靶标？ 3.威尔Tn7 转座破坏正常的谷氨酰胺-果糖-6-磷酸转氨酶 （GFPT）表达？