RETROVIRAL ANTI-SENSE RNA--CELLULAR AND VIRAL RESPONSES

逆转录病毒反义 RNA——细胞和病毒反应

• 批准号: 3191387
• 负责人: EDWARD W HOLMES
• 金额: $ 17.37万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3191387
• 关键词: RNA biosynthesis; Retroviridae; Retroviridae disease; actins; antisense nucleic acid; gene expression; genetic transcription; genetic translation; messenger RNA; nucleic acid probes; nucleic acid sequence; oncogenes; oncogenic virus; polysomes; tissue /cell culture; transposon /insertion element; virus RNA; virus genetics; virus infection mechanism; virus replication

Anti-sense RNA provides a means for specifically blocking the expression of a given gene through formation of a duplex with the target RNA. This technique has also been used to block replication of retroviruses containing RNA complementary to cellular RNA. Thus, this technique offers a potential therapeutic strategy for specifically blocking expression of an oncogenic viral gene product and/or replication of an oncogenic retrovirus. Use of this technology, however, has been limited because the mechanisms by which complementary RNA blocks gene expression or retroviral replication are poorly understood. A better understanding of mechanisms by which anti-sense RNA blocks gene expression and retroviral replication may permit more effective applications of this technology. A model system using retroviral vectors capable of expressing RNA complementary to cellular sequences has been developed. Vectors producing RNA complementary to different regions of the same target mRNA have been isolated. Those vectors producing anti-sense transcripts which are effective in blocking expression of the target mRNA are approximately 100 x more potent than many previously reported, they are complementary to a region of the mRNA which has previously not been considered to be a good target, and they apparently block expression of the complementary mRNA by a mechanism which has not been described previously. The purpose of this proposal is to exploit this model to define the mechanism(s) by which this anti-sense system blocks expression of its compementary cellular mRNA and to determine what effect the complementary cellular RNA has on replication of the retrovirus. Studies are described in which a series of retroviral constructs will be used to 1) define more precisely the regions of the complementary RNAs participating in duplex formation in vivo and in vitro, 2) effects of different RNA transcripts upon duplex formation and stability, and 3) mechanisms by which effective anti-sense constructs block translation of complementary cellular mRNA's. Experiments using this same model are described which will address the following questions: 1) Are cells which produce RNA complementary to that of the retrovirus less susceptible to infection by this retrovirus? If so, what factors affect the extent of this resistance? 2) Do complementary cellular RNAs adversely affect packaging and release of viruses capable of infecting other cells?

反义RNA提供了一种专门阻断 通过与基因形成双链来表达某一特定基因 靶向RNA。这项技术也被用来阻止 复制含有互补的RNA的逆转录病毒 细胞核糖核酸。因此，这项技术提供了一种潜在的治疗方法 特异性阻断致癌病毒表达的策略 致癌逆转录病毒的基因产物和/或复制。使用 然而，这项技术的应用一直受到限制，因为 互补RNA阻断基因表达的机制 或逆转录病毒复制，人们对此知之甚少。更好的 对反义RNA阻断机制的理解 基因表达和逆转录病毒复制可能允许更多 这项技术的有效应用。一个模型系统，使用 能够表达互补的RNA的逆转录病毒载体 细胞序列已经被开发出来。产生RNA的载体 互补于同一靶基因的不同区域 已经被隔离了。那些产生反义的载体 能有效阻断基因表达的转录本 靶基因的效力大约是许多基因的100倍。 此前报道，它们是对一个地区的补充 以前并不被认为是好的 目标，而且它们显然阻止了 通过一种尚未被发现的机制来补充mRNA 如前所述。这项提议的目的是利用 这个模型来定义这个反义的机制(S 系统阻断其组成细胞的mRNA和 为了确定互补的细胞RNA对 逆转录病毒的复制。描述了一项研究，其中 一系列逆转录病毒构建物将用于1)定义更多 准确地说，参与的互补RNA的区域 体内和体外双链形成，2)不同RNA的作用 关于双链形成和稳定性的抄本，以及3) 有效反义构建阻断的机制 互补细胞基因的翻译。实验 使用相同的模型进行了描述，这将解决 以下问题：1)是产生RNA的细胞吗？ 与不易感染的逆转录病毒互补 被这种逆转录病毒感染？如果是这样，是什么因素影响了这一程度 这种抗性？2)对互补的细胞RNA产生不利影响 影响能够感染他人的病毒的包装和释放 细胞？

项目成果

Reversal of creatine kinase translational repression by 3' untranslated sequences.

3非翻译序列逆转肌酸激酶翻译抑制。

• DOI: 10.1126/science.2343304
• 发表时间: 1990
• 期刊: Science (New York, N.Y.)
• 作者: Ch'ng,JL;Shoemaker,DL;Schimmel,P;Holmes,EW
• 通讯作者: Holmes,EW

Antisense RNA complementary to 3' coding and noncoding sequences of creatine kinase is a potent inhibitor of translation in vivo.

与肌酸激酶 3 编码和非编码序列互补的反义 RNA 是体内翻译的有效抑制剂。

• DOI: 10.1073/pnas.86.24.10006
• 发表时间: 1989
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Ch'ng,JL;Mulligan,RC;Schimmel,P;Holmes,EW
• 通讯作者: Holmes,EW