SITE SPECIFIC GENE INSERTION BY TRANSPOSITION

描述（申请人摘要）： 这一提议的核心假设是， 基因在真核细胞染色体中的特异性插入， Tn 7转座蛋白总体目标是 为基因治疗建立生物学基础， 转座子Tn 7蛋白。选择Tn 7是因为它能够整合 转化成DNA序列。初步数据 支持基于Tn 7的插入可以提供安全且 将治疗基因整合到染色体中的有效方法。的 中心假设将在三个具体目标中进行检验：1.将Tn 7 蛋白质转座到真核生物的目标在体外？2.将Tn 7 蛋白质转座到真核生物体内的目标？3.将Tn 7 转座破坏正常谷氨酰胺-果糖-6-磷酸转氨酶 （GFPT）表达？

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?