Forward genetic analysis of Mammalian Resistance to Vira

哺乳动物抗病毒的正向遗传分析

• 批准号: 7202830
• 负责人: BRUCE A BEUTLER
• 金额: $ 39.17万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7202830
• 关键词: Betaherpesvirinae; Herpesviridae disease; capillary electrophoresis; flow cytometry; gene mutation; gene targeting; genetic mapping; genetic susceptibility; genetic transcription; host organism interaction; immune response; immunogenetics; immunomagnetic separation; laboratory mouse; microarray technology; microorganism immunology; positional cloning

Using an unbiased approach (random germline mutagenesis in mice), we have set out to identify host genes that are indispensable for survival early in the course of infection with a large DMA virus (mouse cytomegalovirus; MCMV). In selecting lethality as the endpoint of our screen, we have assured that all targets identified will be essential for defense, without regard to the level at which they function. Approximately 1 in 33 mice born to an ENU-mutagenized sire carries a recessive mutation that is lethal in the context of MCMV infection, suggesting that a large number of genes serve non-redundant functions in host resistance to this pathogen. By screening 12,170 G3 germline mutant mice from 2,028 micropedigrees, we have so far resolved a total of 31 transmissible mutations that markedly impair the innate immune response to MCMV. 8 additional mutations that impair MCMV resistance were identified by cross-screening mice identified in other innate immunity screens, so that 39 mutations have been collected in all, and at the present rate of screening, 25 new mutations should accrue each year. A total of 9 mutations have been identified already, and we have begun to map, assess allelism, and positionally clone those that remain. Informed by parallel efforts in Drosophila (Project 2) and aided by anticipatory resequencing, candidate analysis and positional cloning will be greatly accelerated. Paralogues of mutations that cause MCMV susceptibility will be targeted in mice (Project 3), and using both genetic and biochemical approaches, the mechanisms and pathways utilized in host resistance will be deciphered. Based on comparative estimates of saturation mutagenesis in which lethality is taken as an endpoint, we believe that approximately 11% of all host resistance genes have been struck in our primary screen as performed to date. As a first approximation, offer that about 280 genes comprise the MCMV resistome. The proteins that we identify may fulfill sensing or post-sensing functions. Some may offer very specific protection against MCMV; others may offer protection against many microbes, viral and non-viral alike. Some may fulfill evolutionarily conserved functions (in Drosophila and in mice) while some may be species-specific. We propose to continue the screen, which is the first of its kind in mammalian genetics, and in terms of hit rate, one of the most productive ever described in any model organism. Our goal is to elucidate a large fraction of the proteins that create resistance to viral infection in the mammalian host, and ultimately, to understand how these proteins work.

使用公正的方法（在小鼠中随机种系诱变），我们已着手识别宿主基因 在较大的DMA病毒感染过程中，对于生存而言，这是必不可少的（小鼠 巨细胞病毒; MCMV）。在选择杀伤力作为屏幕的终点时，我们保证 确定的目标对于防御至关重要，而无需考虑其功能的水平。 大约有33只小鼠出生于Enu-Mutagenizatizatization的父亲带有致命的隐性突变 MCMV感染的背景表明，大量基因在 宿主对这种病原体的抗性。通过筛选来自2,028个微鸟类的12,170 G3种系突变小鼠， 到目前为止，我们已经解决了总共31个可传播突变，这些突变显着损害了先天的免疫力 对MCMV的反应。 8个通过盘缩识别的其他损害MCMV电阻的突变 在其他先天免疫筛查中鉴定出的小鼠，因此总共收集了39个突变 目前的筛查率每年应累积25个新突变。总共有9个突变 已经确定了，我们已经开始绘制，评估等位基因，并在定位的地方克隆那些保留的人。 通过果蝇（项目2）的平行努力得出的信息，并通过预期的重新定价， 候选分析和位置克隆将大大加速。引起突变的副产品 MCMV易感性将针对小鼠（项目3），并使用遗传和生化 方法，将在宿主电阻中使用的机制和途径被解密。基于 饱和诱变的比较估计值，其中致死性作为终点，我们认为 所有宿主电阻基因中约有11％在我们的主要屏幕中被击中 日期。作为第一个近似值，提供约280个基因包含MCMV抗性组。蛋白质 我们确定可以实现感应或后感应功能。有些可能会提供非常具体的保护 MCMV;其他人可能会保护许多微生物，包括病毒和非病毒。有些人可能会满足 进化保守的功能（在果蝇和小鼠中），而有些可能是特定物种的。我们 建议继续屏幕，这是哺乳动物遗传学的第一个，就命中率而言， 任何模型生物体中有史以来最有生产力的人之一。我们的目标是阐明很大一部分 在哺乳动物宿主中对病毒感染产生抗性的蛋白质，并最终了解 这些蛋白质如何工作。