DEVELOPMENT OF GENETIC TOOLS FOR TRYPANOSOMA BRUCEI

布氏锥虫遗传工具的开发

• 批准号: 6902322
• 负责人: GEORGE ALAN MARTIN CROSS
• 金额: $ 21.06万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-15 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6902322
• 关键词: RNA interference; Trypanosoma brucei; cell line; gene mutation; gene targeting; genetic library; genetic screening; genetically modified animals; laboratory mouse; protozoal genetics; technology /technique development; transfection /expression vector; trypanosomiasis

DESCRIPTION (provided by the applicant): Human African Trypanosomiasis is returning to levels not seen since the early 20th century. Animal trypanosomiasis remains a significant problem and is a major source of human infection with the virulent Trypanosoma brucei rhodesiense. New drugs are urgently required for all forms of Trypanosomiasis and Leishmaniasis, and understanding the genetic and metabolic circuits of trypanosomes is an important component of the process of validating targets for drug development. Trypanosomes have also attracted general scientific interest as the most widely studied 'differently evolved' eukaryote, with a proven record of contributing important discoveries in membrane biology, cell signaling, telomere biology and gene expression. T. brucei has the potential to continue to contribute to understanding aspects of eukaryotic gene regulation, chromatin modification and telomere structure that are of great contemporary interest. First-generation tools for the genetic manipulation of trypanosomes, pioneered by several laboratories including my own, have severe limitations-of which investigators in the field are increasingly aware-for large-scale analysis of the organism in the context of the soon-to-be-completed genome project and the opportunities that it presents for major advances in understanding the biology of these organisms. A substantial effort is required to develop tools to effectively utilize the genome sequence and to regain momentum in the study of trypanosomes. The aim of this proposal is to improve upon existing cell lines, vectors and protocols, and to develop new approaches, including insertional mutagenesis, for trypanosome genetics. Tools for forward genetics, allowing genome-wide screens by either transposon mutagenesis or the use of RNAi libraries, will become increasingly important to identify the novel genes that undoubtedly regulate the circuitry that is critical to the unique life style, pathogenicity and virulence, of trypanosomes, and which cannot be identified by bioinformatics approaches.

描述（由申请人提供）：非洲人类锥虫病正在恢复到20世纪初以来从未见过的水平。动物锥虫病仍然是一个重大问题，也是人类感染强毒性布氏罗得西亚锥虫的主要来源。目前迫切需要治疗各种形式的锥虫病和利什曼病的新药，而了解锥虫的遗传和代谢回路是药物开发靶点验证过程的重要组成部分。锥虫作为研究最广泛的“不同进化”真核生物也引起了普遍的科学兴趣，在膜生物学、细胞信号传导、端粒生物学和基因表达方面有重要的发现。布鲁氏菌有潜力继续为理解真核基因调控、染色质修饰和端粒结构等方面做出贡献，这些都是当代非常感兴趣的。包括我自己的实验室在内的几个实验室率先开发的第一代锥虫基因操作工具，在即将完成的基因组计划背景下对该生物体进行大规模分析，以及在理解这些生物体的生物学方面取得重大进展的机会方面，存在严重的局限性——该领域的研究人员越来越意识到这一点。需要作出大量的努力来开发工具，以有效地利用基因组序列，并在锥虫的研究中重新获得动力。本建议的目的是改进现有的细胞系、载体和方案，并开发新的方法，包括插入诱变，用于锥虫遗传学。通过转座诱变或使用RNAi文库进行全基因组筛选的前向遗传学工具，对于识别对锥虫独特的生活方式、致病性和毒性至关重要的回路进行调节的新基因将变得越来越重要，而这些基因无法通过生物信息学方法识别。