HOMOLOGOUS RECOMBINATION IN SKIN CELLS FOR GENE THERAPY

描述：（改编自申请人的摘要）-皮肤细胞是 将新基因引入人类的最佳潜在载体之一 用于局部疾病或全身疾病的基因治疗 代谢性疾病迄今为止的工作已经确定了 用于表达和递送的表皮角质形成细胞和成纤维细胞 新基因的局部和系统，但技术和安全问题， 对目前用于引入新的 这些细胞的基因。 这项研究的目的是证明 利用同源重组插入新基因的可行性 表皮角质形成细胞和成纤维细胞中的特异性基因位点。 这 方法应该提供可预测的、持久的和相对安全的 新基因的表达。 该项目的具体目标是：（一） 在β-肌动蛋白中插入一个标记基因，即细菌酶lacZ， 培养的人角质形成细胞和成纤维细胞的位点或进入 角化细胞的角蛋白5基因座和II）选择靶向的 细胞，并评估lacZ表达的持续时间在体外和在 在将靶细胞移植到无胸腺小鼠中后，在体内进行。

DESCRIPTION: (Adapted from the applicant's abstract) - Skin cells are among the best potential vehicles for introducing new genes into humans for the purpose of gene therapy for local disease or for systemic metabolic disease. Work to date has established the usefulness of epidermal keratinocytes and fibroblasts for expressing and delivering new genes locally and systemically, but technical and safety issues raise questions about currently employed methods used to introduce new genes into those cells. The goal of this research is to demonstrate the feasibility of using homologous recombination to insert new genes into specific gene loci in epidermal keratinocytes and fibroblasts. This method should provide predictable, durable, and relatively safe expression of new genes. The specific aims of this project are I) to insert a marker gene, the bacterial enzyme lacZ, into the beta-actin locus of cultured human keratinocytes and fibroblasts or into the keratin 5 locus of keratinocytes and II) to select clones of targeted cells and to evaluate the duration of lacZ expression in vitro and in vivo, following transplantation of the targeted cells into athymic mice.