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

 描述(申请人提供)：数以千计的基因组已经被测序，但它们编码的大多数基因的功能在很大程度上仍然未知。我的实验室旨在通过开发广泛适用的工具，极大地加快对未定性基因的研究，来改变我们设计生物学的能力。在这个新的创新者项目中，我们将使用化学遗传学为绿藻衣藻中数以千计的未知基因分配功能，衣藻衣藻是一个强大的模型系统，主要研究纤毛的生物发生和运动性、中心体、光合作用、电子传输、细胞器间通信和光遗传学。我们将利用我的实验室开发的改变游戏规则的工具，包括这种有机体中第一个全基因组范围的索引突变集合，以及跟踪10万个突变体库中突变丰度的方法。我们将量化代表基因组中几乎所有基因的突变体在200种生长条件下的生长缺陷和增强。此外，在专注于纤毛功能的推力中，我们将在50个最近发现的纤毛功能小分子调节剂存在的情况下，量化所有突变体的纤毛表型。我们将使用这些数据来准确预测数千个未确定特征的基因的功能，这一原理是基于相同途径中功能的基因突变通常表现出相似的表型。此外，我们将利用化学-遗传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，重点放在纤毛生物发生所必需的新基因上。更广泛地说，该项目将作为一个平台，推动尖端工具的开发，以便系统和定量地探索整个生命树的基因功能。

 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.