HAMMERHEAD RIBOZYME TRAGETED TO MURINE LEUKEMIA VIRUS

小鼠白血病病毒(MuLV)是从SO野生型小鼠中分离到的。 加利福尼亚。这些非急性白血病病毒属于 C逆转录病毒。他们没有任何已知的致癌基因，也不能 在培养中转化细胞。然而，它们会在野外引起脾淋巴瘤， 以及敏感的近交系小鼠。这些野鼠MuLV有很多 与人类T细胞白血病病毒相似。作为人类的典范 淋巴瘤/白血病，野生型小鼠MuLV诱导的脾淋巴瘤 类似于儿童急性淋巴细胞性白血病。总的目标是 目前的研究是开发一种抗逆转录病毒的方法 会导致病毒RNA的特定裂解，抑制病毒感染 在培养细胞和病毒诱导的小鼠脾淋巴瘤中。 最近发现的核酶表明，特定的RNA分子可以 以酶的形式工作，降解目标RNA分子。在各种不同的 核酶，锤头状核酶的催化结构域似乎是 最简单的一种，催化中心可以很容易地结合到 反义RNA。针对特定RNA的核酶的特异性靶向 然而，细胞内部似乎构成了一个主要问题。在建议的 研究，将设计、合成和靶向锤头状核酶 通过利用psi序列(逆转录病毒)来对抗MuLV RNA RNA包装信号)为基础的逆转录病毒载体。这是意料之中的 这样的序列将使核酶和MuLV基因组RNA共存 在感染细胞的细胞质微环境中，帮助反式- 病毒核糖核酸的裂解。此外，这样的核酶构建物将 也被整合到小鼠的生殖系中。转基因生物的保护 小鼠抗病毒攻击和小鼠轻度感染小鼠MuLV 10A1 因此，淋巴肿大将为延长这一期限提供合理的基础。 逆转录病毒相关人类癌症的治疗策略 细胞白血病和逆转录病毒引起的获得性等相关疾病 免疫缺陷综合征(艾滋病)

Murine leukemia viruses (MuLV) have been isolated from the wild mice of So. California. These non-acute leukemia viruses belong to the family of type C retroviruses. They do not possess any known oncogene and are unable to transform cells in culture. However, they cause splenic lymphoma in wild, as well as, susceptible inbred mice. These wild mouse MuLVs have many similarities with human T-cell leukemia Virus. As a model for human lymphoma/leukemia, the wild mouse MuLV-induced splenic lymphoma closely resembles childhood acute lymphoblastic leukemia. The overall objective of the present investigation is to develop an anti-retroviral approach which will cause specific cleavage of the viral RNA, inhibiting virus infection in a cultured cell and virus-induced splenic lymphoma in mice. The recent discovery of ribozymes has shown that specific RNA molecules can work as enzymes and degrade target RNA molecules. Of t he various ribozymes, the hammerhead ribozyme catalytic domain appears to be the simplest one, and the catalytic center can be easily incorporated into anti-sense RNA. Specific targeting of a ribozyme against a particular RNA inside the cell, however, appears to pose a major problem. In the proposed study, a hammer-head ribozyme will be designed, synthesized, and targeted against the MuLV RNA by taking advantage of the psi sequence (retroviral RNA packaging signal) of an MuLV-based retroviral vector. It is expected that such a sequence will colocalize the ribozyme and the MuLV genomic RNA in the cytoplasmic micro-environment of the infected cell, aiding trans- cleavage of the viral RNA. Additionally, such a ribozyme construct will also be incorporated in the germline of mice. Protection of the transgenic mice against virus challenge and mild mouse MuLV 10A1-induced lymphomagenesis would thus provide a rational basis for extending this strategy for the treatment of retrovirus-associated human cancer such as T- cell leukemia and retrovirus induced allied diseases like acquired immunodeficiency syndrome (AIDS)