Reconstructing the Global Epistasis Network for Aging

重建全球老龄化上位网络

• 批准号: 10396088
• 负责人: HAO LI
• 金额: $ 20.19万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10396088
• 关键词: Address; Aging; Animal Model; Bioinformatics; Biological Assay; Biology of Aging; CRISPR interference; Cell Count; Cell division; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Computer Analysis; DNA Sequence; Data; Data Analyses; Daughter; Engineering; Eukaryota; Foundations; Gene Structure; Genes; Genetic; Genetic Engineering; Genetic Epistasis; Genomics; High-Throughput Nucleotide Sequencing; Libraries; Light; Liquid substance; Longevity; Maps; Measures; Mediating; Microdissection; Molecular Genetics; Mothers; Nature; Pathway interactions; Persons; Phenotype; Regulation; Research; Research Project Summaries; Saccharomycetales; Techniques; Technology; Testing; Validation; Yeasts; base; combinatorial; daughter cell; gene network; genetic analysis; genetic strain; genome-wide; high throughput analysis; insight; longevity gene; mutant; new technology; novel; programs; reconstruction

Project Summary Research in the biology of aging has seen rapid advance since the introduction of molecular genetics to the field. In the past few decades, genetic analyses of model organisms have led to the discovery of a number of genes/pathways that regulate lifespan, leading to important insight into the mechanisms of aging. However a global picture of the gene network that regulates lifespan is still elusive. To classify longevity mutants based on their mechanisms of action, to place genes influencing lifespan into pathways, and to delineate the downstream effectors of the lifespan extension for the longevity mutants, it is necessary to systematically analyze the epistatic relations between genes for the lifespan phenotype. So far this has not been possible in any model organism due to the sheer number of double mutants that need to be analyzed and the low throughput nature of the traditional lifespan assays. Here we propose to systematically reconstruct the global epistasis network for aging using budding yeast as the model organism and replicative lifespan as the phenotype. We will develop a high throughput approach to generate all possible single and double mutants and measure their lifespan simultaneously (Aim 1). Our approach will be based on a number of new technologies: 1) a novel genetically engineered strain that makes it possible to measure lifespan based on cell counting in liquid culture; 2) CRISPR/dCas9 based technology to generate a pooled library of single and double mutants, each carrying a unique DNA sequence identifier; 3) high throughput sequencing to count the number of cells of the pooled mutants simultaneously. We will use the comprehensive lifespan data to reconstruct the global epistasis map through computational analysis. We will also test some of the discovered epistatic relations both in yeast and in worms (Aim 2). We expect that this project will produce unprecedented and comprehensive data on the epistasis network that controls lifespan in a canonical model organism. The analysis of this data will lead to important insights into the functional organization of genes in the context of aging and mechanisms for the lifespan extension in long-lived mutants. Such insights might be transferrable to higher eukaryotes, in which a systematic reconstruction of the epistasis network for aging is not feasible.

项目摘要 自分子遗传学引入以来，衰老生物学研究取得了迅速进展 领域在过去的几十年里，对模式生物的遗传分析已经导致发现了许多 基因/调节寿命的途径，导致对衰老机制的重要见解。然而 调节寿命的基因网络的全球图景仍然难以捉摸。对长寿突变体进行分类 它们的作用机制，将影响寿命的基因置于通路中，并描绘出 对于长寿突变体的寿命延长的下游效应子，有必要系统地 分析寿命表型基因间的上位关系。到目前为止， 由于需要分析的双突变体的绝对数量和低的 传统寿命测定的通量性质。 在这里，我们建议系统地重建全球上位网络老化使用芽殖酵母作为 模式生物和复制寿命作为表型。我们将开发一种高通量方法， 产生所有可能的单突变体和双突变体，并同时测量它们的寿命（目标1）。我们 这种方法将基于一些新技术：1）一种新的基因工程菌株， 可以基于液体培养中的细胞计数来测量寿命; 2）基于CRISPR/dCas 9的技术， 产生单突变体和双突变体的合并文库，每个突变体携带独特的DNA序列标识符; 3） 高通量测序以同时计数合并的突变体的细胞数。我们将使用 综合寿命数据，通过计算分析重建全局上位性图。我们将 还测试了酵母和蠕虫中发现的一些上位关系（目标2）。 我们希望这个项目将产生前所未有的关于上位网络的全面数据， 控制着典型模式生物的寿命对这些数据的分析将导致对 衰老背景下基因的功能组织以及长寿者寿命延长的机制 变种人这样的见解可能可以转移到高等真核生物，其中系统地重建了 衰老的上位性网络是不可行的。

项目成果

In Vivo Dopamine Neuron Imaging-Based Small Molecule Screen Identifies Novel Neuroprotective Compounds and Targets.

基于体内多巴胺神经元成像的小分子筛选可识别新型的神经保护化合物和靶标。

• DOI: 10.3389/fphar.2022.837756
• 发表时间: 2022
• 期刊: Frontiers in pharmacology
• 影响因子: 5.6
• 作者: Kim GJ;Mo H;Liu H;Okorie M;Chen S;Zheng J;Li H;Arkin M;Huang B;Guo S
• 通讯作者: Guo S

A zebrafish screen reveals Renin-angiotensin system inhibitors as neuroprotective via mitochondrial restoration in dopamine neurons.

• DOI: 10.7554/elife.69795
• 发表时间: 2021-09-22
• 期刊: eLife
• 影响因子: 7.7
• 作者: Kim GJ;Mo H;Liu H;Wu Z;Chen S;Zheng J;Zhao X;Nucum D;Shortland J;Peng L;Elepano M;Tang B;Olson S;Paras N;Li H;Renslo AR;Arkin MR;Huang B;Lu B;Sirota M;Guo S
• 通讯作者: Guo S

Further Extension of Lifespan by Unc-43/CaMKII and Egl-8/PLCβ Mutations in Germline-Deficient Caenorhabditis elegans.

• DOI: 10.3390/cells11223527
• 发表时间: 2022-11-08
• 期刊: Cells
• 影响因子: 6