Reverse genetics of Chlamydomonas using TILLING

使用 TILLING 进行衣藻反向遗传学

基本信息

批准号：
7280452
负责人：
KRISHNA K NIYOGI
金额：
$ 29.64万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-01 至 2009-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7280452
关键词：
Animal Model Antioxidants Appendix Arabidopsis Biochemistry Biogenesis Biological Biological Process Biology Cell-Cell Adhesion Cellular biology Chemicals Chlamydomonas Chlamydomonas reinhardtii Cilia Collaborations Communities Complementary DNA DNA Sequence Data Databases Deposition Detection Development Diploidy Disease Drosophila genus Dyskinetic syndrome Eukaryota Eukaryotic Cell Evolution Farming environment Flagella Frequencies Gases Gene Targeting Generations Genes Genetic Genetic Crosses Genome Germ Cells Goals Green Algae Haploidy Homologous Gene Individual Insertional Mutagenesis Institutes Joints Juglans Knock-out Large-Scale Sequencing Letters Libraries Maize Methods Modeling Molecular Mutagenesis Mutation Nuclear Organism Participant Personal Satisfaction Phenotype Photosynthesis Phylogenetic Analysis Physiology Plants Plastids Point Mutation Polycystic Kidney Diseases Population Process RNA Interference Regulation Research Research Personnel Resources Screening procedure Sequence Analysis Series Services Sex Chromosomes Signal Transduction Single Nucleotide Polymorphism Structure Study models Technology Work Zebrafish cost density functional genomics gene function genetic resource genome sequencing homologous recombination human disease indexing insertion/deletion mutation insight interest kinetosome knockout gene loss of function mutation member mutant positional cloning programs tool vitamin metabolism

项目摘要

DESCRIPTION (provided by applicant): Project Summary: The unicellular green alga Chlamydomonas reinhardtii is the model organism of choice for studying many fundamental aspects of eukaryotic cell biology, physiology, and biochemistry. As examples, the biogenesis and function of flagella/cilia, basal bodies, and plastids are most easily investigated using Chlamydomonas. A draft sequence of the Chlamydomonas nuclear genome at >9X coverage has been released by the DOE's Joint Genome Institute, but the lack of reverse genetic resources is currently a major limitation for functional genomics of Chlamydomonas. TILLING is a high-throughput reverse genetics approach that combines traditional mutagenesis with modern technology for detection of point mutations. TILLING can provide an allelic series of mutations in any gene of interest. The goal of this project is to establish a publicly available TILLING resource for the Chlamydomonas research community, in collaboration with the Seattle TILLING Project, which has successfully developed similar resources for the Arabidopsis, Drosophila, and zebrafish communities. The specific aims of this proposal are (1) to optimize UV/chemical mutagenesis of haploid Chlamydomonas and generate mutant populations for TILLING, (2) to perform pilot TILLING projects to determine if the haploid mutation density is sufficient for cost-effective TILLING, (3) to optimize UV/chemical mutagenesis of diploid Chlamydomonas and generate mutant populations for TILLING, (4) to perform pilot TILLING projects using a mutagenized diploid population, and (5) to provide a TILLING service to the Chlamydomonas research community. There is broad community support for the development of a TILLING resource for Chlamydomonas. The mutant populations and data generated by this project will be publicly available from the Chlamydomonas stock center and database. Relevance: Chlamydomonas is an important model for basic biological mechanisms and for human disease processes such as polycystic kidney disease, primary cilia dyskinesia, and Rh null disease. This project will generate a resource that will enable the determination of gene function for many genes that have been discovered by sequencing the Chlamydomonas genome.

描述（由申请人提供）：项目摘要：单细胞绿藻莱茵衣藻是研究真核细胞生物学、生理学和生物化学许多基本方面的首选模式生物。例如，使用衣藻最容易研究鞭毛/纤毛、基体和质体的生物发生和功能。美国能源部联合基因组研究所已发布了覆盖率 >9X 的衣藻核基因组草图序列，但反向遗传资源的缺乏目前是衣藻功能基因组学的主要限制。 TILLING 是一种高通量反向遗传学方法，它将传统诱变与现代点突变检测技术相结合。 TILLING 可以在任何感兴趣的基因中提供一系列等位基因突变。该项目的目标是与西雅图 TILLING 项目合作，为衣藻研究界建立一个公开可用的 TILLING 资源，该项目已成功为拟南芥、果蝇和斑马鱼社区开发了类似的资源。该提案的具体目标是 (1) 优化单倍体衣藻的紫外线/化学诱变并生成 TILLING 突变群体，(2) 进行试点 TILLING 项目以确定单倍体突变密度是否足以进行具有成本效益的 TILLING，（ 3) 优化二倍体衣藻的紫外线/化学诱变并产生 TILLING 突变群体，(4) 使用诱变的二倍体群体进行试点 TILLING 项目， (5) 为衣藻研究界提供 TILLING 服务。衣藻 TILLING 资源的开发得到了广泛的社区支持。该项目产生的突变体种群和数据将在衣藻库存中心和数据库中公开提供。相关性：衣藻是基本生物学机制和人类疾病过程（如多囊肾病、原发性纤毛运动障碍和 Rh 缺失病）的重要模型。该项目将生成一种资源，能够确定通过衣藻基因组测序发现的许多基因的基因功能。