ENHANCEMENT AND MODULATION OF ANTI-SENSE RNA ACTIVITY

反义 RNA 活性的增强和调节

• 批准号: 3191378
• 负责人: JONATHAN G IZANT
• 金额: $ 10.11万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3191378
• 关键词: R factors; RNA; Xenopus; cell transformation; egg /ovum; gene expression; genetic manipulation; genetic promoter element; genetic transcription; genetic translation; germ layers; messenger RNA; molecular cloning; nucleic acid hybridization; ribosomal RNA; structural genes; transfer RNA

Antisense RNA transcribed from "flipped gene" plasmids is an effective method for cell autonomous suppression of exogenous (e.g. viral) as well as endogenous gene activity. We propose to expand our research program on anti-sense RNA to include two additional facets which will encompass RFA #87-CA-18 "Studies of Functional Anti-sense RNA in Oncogenic Viral Systems". Anti-sense gene chimeras will be constructed with URNA, tRNA, and rRNA genes. The level of transcription and the anti-sense inhibition activity of the hybrid genes will be tested in Xenopus oocytes and cultured mammalian cells. The goal of these studies is to exploit both the extremely active promoter signals in these structural RNA genes and, if possible, harness some of the special biological activity of the URNA, tRNA, and rRNA genes to enhance anti-sense RNA activity. Factors that modulate anti-sense RNA activity will be assessed in cultured mammalian cells and in Amphibian embryos. Somatic cell lines with high and low levels of RNA duplex destablizing activity will be selected and the nature of the "unwindase" characterized. Xenopus embryo germ layers and Xenopus tadpole tissues will be tested for unwindase activity that might contribute to tissue- specific gene expression. The capacity of the aforementioned chimeric anti-sense RNAs to overcome the unwindase block will be assessed in each system. Our goal is both to understand and to circumvent this crucial aspect of in vivo RNA-RNA interaction.

从“翻转基因”质粒转录的反义RNA是一种 用于细胞自主抑制外源性（例如， 病毒）以及内源性基因活性。 我们建议扩大 我们的反义RNA研究计划包括两个额外的 将涵盖RFA #87-CA-18“功能研究”的方面 在致癌病毒系统中的反义RNA”。 反义基因嵌合体将用URNA、tRNA和 rRNA基因。 转录水平和反义 将在非洲爪蟾中测试杂交基因的抑制活性 卵母细胞和培养的哺乳动物细胞。 这些研究的目的 是利用这些基因中极其活跃的启动子信号， 结构RNA基因，如果可能的话，利用一些特殊的 URNA、tRNA和rRNA基因的生物活性增强 反义RNA活性。 调节反义RNA活性的因素将在 培养的哺乳动物细胞和两栖动物胚胎。 体细胞 具有高水平和低水平RNA双链体去稳定化活性的品系 将被选择和性质的“unwindase”的特点。 非洲爪蟾胚胎胚层和非洲爪蟾蝌蚪组织将 测试解旋酶的活性，可能有助于组织- 特异性基因表达 上述能力 克服解旋酶阻断的嵌合反义RNA将被 在每个系统中评估。 我们的目标是了解和 绕过了体内RNA-RNA相互作用的这一关键方面。