SITE SPECIFIC GENE INSERTION BY TRANSPOSITION

描述(申请人摘要)： 这一提议的中心假设是位置和序列 可以将基因特异性插入真核细胞的染色体中 不含Tn7转座蛋白。总体目标是 基于对基因治疗的使用为基因治疗建立生物学基础 转座子Tn7蛋白。TN7之所以被选中，是因为它能够集成 转化成具有特异性和效率的DNA序列。初步数据 支持一种假设，即基于Tn7的插入可以提供安全和 将治疗性基因整合到染色体中的有效方法。这个 中心假说将在三个具体目标中得到检验：1.Tn7 体外转座到真核细胞靶点的蛋白质？2.Tn7 蛋白质在体内转位成真核靶点？3.Tn7 转座干扰正常的谷氨酰胺-果糖-6-磷酸转氨酶 (GFPT)表达式？

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?