TARGETED TRANSGENESIS FOR EPIDERMAL GENE THERAPY

表皮基因治疗的靶向转基因

基本信息

批准号：
09670893
负责人：
YAMANISHI Kiyofumi
金额：
$ 1.98万
依托单位：
KYOTO PREFECTURAL UNIVERSITY OF MEDICINE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670893/
关键词：
GENE TARGETING TRANSGENIC MICE TRANSGLUTAMINASE GENE THERAPY トランスグルタミナーゼ1

项目摘要

We generated transgenic mice expressing the Escherichia coli beta-galactosidase gene (lalphaZ) directed by the 5' upstream region of human transglutaminase 1 (TGase 1) gene. beta-Galactosidase histochemistry revealed that the TGMI-lalphaZ transgene was expressed in terminally differentiating keratinocytes of the upper layers of the stratified squamous epithelia in embryonic, neonatal and adult transgenic mice. The expression pattern was similar to that of endogenous TGase 1 niRNA detected by in situ hybridization. Furthermore, topical application of a phorbol ester on the adult tail skin enhanced expression of the transgene as well as TGase 1 mRNA in the epidermis. Thus, the 2.5-kb 5' upstream sequence of the human TGase 1 gene may be useful for epidermis and terminal differentiation-specific gene therapy in the stratified squamous epithelia.Next, we generated mice lacking the TGase 1 gene, and showed that they have erythrodermic skin with abnormal keratinization. In their stratum corneum, degradation of nuclei and keratohy aim F-granules was incomplete and cell envelope assembly was defective. The skin barrier function of TGase 1-null mice was markedly impaired, and these mice died within 4-5 h after birth. These results clearly demonstrate that the TGase 1 gene is essential to the development and maturation of the stratum corneum and to adaptation to the environment after birth. Thus, these TGase 1 knock-out mice may be a useful model for severe cases of LI gene therapy.

我们产生了表达大肠杆菌β-半乳糖苷酶基因（Lalphaz）的转基因小鼠，该基因的5'上游区域（tgase 1）基因指向5'上游区域。 β-半乳糖苷酶的组织化学表明，TGMI-Lalphaz转基因在胚胎，新生儿和成年转基因小鼠的分层鳞状上皮的上层的终末角质形成细胞中表达。表达模式与原位杂交检测到的内源性TGase 1 NiRNA相似。此外，佛比尔酯在成年尾皮上的局部应用增强了转基因的表达以及表皮中的TGase 1 mRNA。因此，人类TGase 1基因的2.5-kb 5'上游序列在分层的鳞状上的鳞状上的表皮和末端分化特异性基因疗法可能很有用。我们产生了缺乏TGase 1基因的小鼠，并表明它们具有异常皮肤具有异常呈异常。在角膜层中，核和角膜纤维颗粒的降解不完整，细胞包络组装有缺陷。 TGASE 1-NULL小鼠的皮肤屏障功能显着受损，这些小鼠在出生后4-5小时内死亡。这些结果清楚地表明，TGase 1基因对于角膜层的发展和成熟至关重要，并在出生后适应环境至关重要。因此，这些TGASE 1敲除小鼠可能是严重的LI基因治疗病例的有用模型。