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（由申请人提供）：数千个基因组已被测序，但它们编码的大多数基因的功能仍然很大程度上未知。我的实验室旨在通过广泛适用的工具来转变我们设计生物学的能力，这些工具极大地加速了生物学的研究。在这个新创新者项目中，我们将利用化学遗传学为绿藻莱茵衣藻中数千个未表征的基因分配功能，这是一个强大的模型系统，对于纤毛生物发生和研究至关重要。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用于跟踪突变体丰度的方法。我们将量化代表 200 种生长条件下基因组中几乎所有基因的 100,000 个突变体的生长缺陷和增强。在 50 种最近发现的纤毛功能小分子调节剂存在下，对所有突变体进行定量纤毛表型，我们将根据在同一途径中发挥作用的基因突变体通常表现出的原理，使用这些数据准确预测数千个未表征基因的功能。此外，我们将利用化学-遗传相互作用来鉴定纤毛功能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，重点关注纤毛所必需的新基因。更广泛地说，该项目将作为推进尖端工具开发的平台，以系统地、定量地探测整个生命树的基因功能。