• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>BMC Biotechnology >Generation of a tumor- and tissue-specific episomal non-viral vector system
【24h】

Generation of a tumor- and tissue-specific episomal non-viral vector system

机译：肿瘤和组织特异性游离型非病毒载体系统的产生

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Background A key issue for safe and reproducible gene therapy approaches is the autologous and tissue-specific expression of transgenes. Tissue-specific expression in vivo is either achieved by transfer vectors that deliver the gene of interest into a distinct cell type or by use of tissue-specific expression cassettes. Here we present the generation of non-viral, episomally replicating vectors that are able to replicate in a tissue specific manner thus allowing tissue specific transgene expression in combination with episomal replication. The episomal replication of the prototype vector pEPI-1 and its derivatives depends exclusively on a transcription unit starting from a constitutively active promoter extending into the scaffold/matrix attachment region (S/MAR). Results Here, we exchanged the constitutive promoter in the pEPI derivative pEPito by the tumor specific alpha fetoprotein (AFP) or the muscle specific smooth muscle 22 (SM22) promoter leading to specific transgene expression in AFP positive human hepatocellular carcinoma (HUH7) and in a SM22 positive cell line, respectively. The incorporation of the hCMV enhancer element into the expression cassette further boosted the expression levels with both promoters. Tissue specific-replication could be exemplary proven for the smooth muscle protein 22 (SM22) promoter in vitro. With the AFP promoter-driven pEPito vector hepatocellular carcinoma-specific expression could be achieved in vivo after systemic vector application together with polyethylenimine as transfection enhancer. Conclusions In this study we present an episomal plasmid system designed for tissue specific transgene expression and replication. The human AFP-promoter in combination with the hCMV enhancer element was demonstrated to be a valuable tissue-specific promoter for targeting hepatocellular carcinomas with non-viral gene delivery system, and tissue specific replication could be shown in vitro with the muscle specific SM22 promoter. In combination with appropriate delivery systems, the tissue specific pEPito vector system will allow higher tissue-specificity with less undesired side effects and is suitable for long term transgene expression in vivo within gene therapeutical approaches.
机译：背景技术安全且可重复的基因治疗方法的关键问题是转基因的自体和组织特异性表达。体内组织特异性表达可以通过将目的基因传递到不同细胞类型的转移载体来实现，也可以通过使用组织特异性表达盒来实现。在这里，我们介绍了能够以组织特异性方式复制的非病毒，游离型复制载体的产生，从而使组织特异性转基因表达与游离型复制相结合。原型载体pEPI-1及其衍生物的游离复制完全取决于转录单元，该转录单元从组成型活性启动子开始，延伸至支架/基质附着区（S / MAR）。结果在这里，我们通过肿瘤特异性α胎蛋白（AFP）或肌肉特异性平滑肌22（SM22）启动子交换了pEPI衍生物pEPito中的组成型启动子，导致在AFP阳性人肝细胞癌（HUH7）和肝癌中特异性转基因表达。分别为SM22阳性细胞系。将hCMV增强子元件掺入表达盒进一步提高了两个启动子的表达水平。组织特异性复制可以在体外被平滑肌蛋白22（SM22）启动子证实。使用AFP启动子驱动的pEPito载体，与聚乙烯亚胺作为转染增强剂一起应用全身载体后，可在体内实现肝细胞癌特异性表达。结论在这项研究中，我们提出了一种用于组织特异性转基因表达和复制的游离型质粒系统。人类AFP启动子与hCMV增强子的结合被证明是有价值的组织特异性启动子，可通过非病毒基因递送系统靶向肝细胞癌，并且组织特异性复制可通过肌肉特异性SM22启动子在体外显示。结合适当的递送系统，组织特异性pEPito载体系统将具有更高的组织特异性，副作用更少，并且适用于基因治疗方法中的体内长期转基因表达。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号