ENHANCEMENT AND MODULATION OF ANTI-SENSE RNA ACTIVITY

反义 RNA 活性的增强和调节

• 批准号: 3191379
• 负责人: JONATHAN G IZANT
• 金额: $ 10.15万
• 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 1991-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3191379
• 关键词: R factors; RNA; RNA splicing; Xenopus; antisense nucleic acid; 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