Transforming our understanding of eukaryotic gene functions through chemical genetics in the green alga Chlamydomonas reinhardtii

通过绿藻莱茵衣藻的化学遗传学改变我们对真核基因功能的理解

• 批准号: 8955354
• 负责人: Martin Casimir Jonikas
• 金额: $ 60.42万
• 依托单位: CARNEGIE INSTITUTION OF WASHINGTON, D.C.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8955354
• 关键词: Advanced Development; Biogenesis; Biological Models; Biology; Centrosome; Chlamydomonas; Chlamydomonas reinhardtii; Cilia; Collaborations; Collection; Communication; Communities; Data; Defect; Disease; Electron Transport; Engineering; FDA approved; Functional disorder; Genes; Genetic; Genome; Germ Cells; Green Algae; Growth; Hereditary Disease; Human; Human body; Label; Life; Malignant Neoplasms; Methods; Organelles; Organism; Pathway interactions; Phenotype; Photosynthesis; Play; Polycystic Kidney Diseases; Process; Role; Trees; base; cell motility; chemical genetics; ciliopathy; cilium biogenesis; congenital heart disorder; drug market; gene function; genome-wide; indexing; insight; mutant; novel; optogenetics; public health relevance; small molecule; tool

 DESCRIPTION (provided by applicant): Thousands of genomes have been sequenced, but the functions of most of the genes that they encode remain largely unknown. My lab aims to transform our ability to engineer biology by developing broadly applicable tools that dramatically accelerate the study of uncharacterized genes. In this New Innovator project, we will use chemical genetics to assign functions to thousands of uncharacterized genes in the green alga Chlamydomonas reinhardtii, a powerful model system central to studies of ciliary biogenesis and motility, centrosomes, photosynthesis, electron transport, inter-organelle communication and optogenetics. We will leverage game-changing tools developed by my lab, including the first genome-wide collection of indexed mutants in this organism and methods for tracking mutant abundances in pools of 100,000 mutants. We will quantify the growth defects and enhancements of mutants representing nearly all genes in the genome across 200 growth conditions. Additionally, in a thrust focusing on ciliary function, we will quantify ciliary phenotypes for all mutants in the presence of 50 recently discovered small molecule modulators of cilia function. We will use these data to accurately predict functions for thousands of uncharacterized genes based on the principle that mutants in genes that function in the same pathway usually show similar phenotypes. Furthermore, we will identify the genetic targets of the small molecule modulators of cilia function using chemical-genetic interactions. We will dissect gene functions in collaboration with the scientific community, with a focus on novel genes essential for cilia biogenesis. More broadly, the project will serve as a platform for advancing the development of cutting-edge tools to systematically and quantitatively probe gene functions throughout the tree of life.

 描述（由适用提供）：已经测序了成千上万的基因组，但是它们编码的大多数基因的功能仍然在很大程度上未知。我的实验室旨在通过开发广泛适用的工具来改变我们的生物学能力，从而大幅度地加速未表征基因的研究。在这个新的创新项目中，我们将使用化学遗传学将功能分配给绿色藻类衣原体Reinhardtii中的数千个未表征的基因，这是一种强大的模型系统，是纤毛生物发生和运动性研究，中心体，光合作用，光合作用，电子运输，电子运输，跨层次交流和选择性的核心系统的核心系统。我们将利用我的实验室开发的改变游戏规则的工具，包括该生物体中第一个全基因组索引突变体的集合以及跟踪100,000个突变体池中突变体丰度的方法。我们将量化代表200个生长条件的基因组几乎所有基因的突变体的生长缺陷和增强。此外，在关注睫状功能的推力中，我们将在50个最近发现的纤毛功能的小分子调节剂存在下，对所有突变体的睫状表型进行量化。我们将使用这些数据来准确地预测数千个未表征基因的功能，即基因在同一途径中起作用的基因中的突变体通常显示出相似的表型。此外，我们将使用化学基因相互作用来确定纤毛功能的小分子调节剂的遗传靶标。我们将与科学界合作剖析基因功能，重点是纤毛生物发生必不可少的新基因。从更广泛的角度来看，该项目将成为一个平台，以促进在整个生命之树中进行系统和定量探测基因功能的开发。