HAIRPIN RIBOZYME GENE THERAPY FOR HCV INFECTION

描述：(改编自调查人员摘要)此第二阶段SBIR 建议发展发夹状核酶(RZ)基因治疗 丙型肝炎病毒感染的治疗。丙型肝炎病毒是引起 非甲非乙型肝炎，这是一个严重的全球卫生问题，以及 核酶是可以被改造为切割和失活的rna分子。 其他RNA分子以特定于序列的方式。调查人员认为 丙型肝炎病毒特别适合于核酶介导的治疗，因为病毒 基因组只以RNA的形式存在，因为丙型肝炎病毒只编码 一份主要成绩单。因此，申请人的理由是任何乳沟 这个RNA中的靶序列应该会取消病毒的表达。伊姆索尔有 产生了两个针对丙型肝炎病毒正链RNA的发夹状核酶，并在 确定了该项目的第一阶段组件--其他Rz 负链，由于其低表达而成为更具吸引力的靶子 水平。在这项第二阶段的建议中，首席调查员建议 前六位Rz的催化活性和稳定性的优化。 将使用肝细胞特异性启动子将Rz表达为单一或 多顺反子转录本及其多核酶载体的有效性 将在组织培养报告模型中进行测试。丙型肝炎病毒的研制 提出了肝细胞培养中的复制系统，用于最终测试 在大鼠毒性研究中进行临床前安全性评估前的核酶载体。 建议的商业应用：不可用

DESCRIPTION: (Adapted from the investigator's abstract) This Phase II SBIR proposal is aimed at the development of hairpin Ribozyme (RZ) gene therapy for the treatment of Hepatitis C virus (HCV) infection. HCV is the major cause of non-A, non-B hepatitis, which is a serious worldwide health problem, and ribozymes are RNA molecules that can be engineered to cleave and inactivate other RNA molecules in a sequence-specific fashion. The investigators believe that HCV is especially suited for ribozyme-mediated therapy, because the viral genome is present exclusively in the form of RNA and because HCV encodes only one primary transcript. Therefore, the applicants reason that cleavage of any target sequence within this RNA should abolish virus expression. Immusol has generated two hairpin ribozymes targeting the HCV positive strand RNA, and in the Phase I component of this project, additional Rz were identified targeting the negative strand, an even more attractive target due to its low expression level. In this Phase II proposal, the Principal Investigator proposes the optimization of the catalytic activity and stability of the top six Rz. Hepatocyte specific promoters will be used to express the Rz as single or polycistronic transcripts and the effectiveness of these multi-ribozyme vectors will be tested in a tissue culture reporter model. Development of an HCV replication system in liver cell culture is proposed, for final testing of the ribozyme vector before pre-clinical safety evaluation in a rat toxicity study. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE