Genetic Recombination in C. elegans

线虫中的基因重组

基本信息

批准号：
9177624
负责人：
ANNE M VILLENEUVE
金额：
$ 41.35万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-06-01 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9177624
关键词：
ATAC-seq Address Affect Aneuploidy Architecture Biochemical Caenorhabditis elegans Cell Count Cell division Cells ChIP-seq Chromatin Chromosome Structures Chromosomes Congenital Abnormality Coupling Cytolysis DNA DNA Damage DNA Double Strand Break DNA Repair Development Diploidy Ensure Environment Equilibrium Event Failure Generations Genes Genetic Genetic Recombination Genetic Screening Genome Genomic approach Germ Cells Goals Haploidy Health Homologous Gene Human Image Immunofluorescence Immunologic Individual Licensing Life Mechanics Meiosis Meiotic Recombination Modeling Mutagenesis Nematoda Nuclear Organism PLK1 gene Pathway interactions Play Ploidies Procedures Process Property Prophase Proteins Regulation Repair Complex Research Resolution Role Site Spontaneous abortion Synaptonemal Complex System Testing Work base density design genetic analysis genetic approach genome integrity genome-wide homologous recombination in vivo mutant programs protein complex repaired success tumor progression

项目摘要

Project Summary: Our long term goal is to elucidate how genetic recombination contributes to the faithful inheritance of chromosomes during meiosis, the specialized cell division program by which diploid organisms generate haploid gametes. Chromosome inheritance during meiosis relies on the formation of double-strand DNA breaks (DSBs) and repair of a subset of these DSBs as inter-homolog crossovers (COs). Failure to form COs leads to chromosome missegregation and consequent aneuploidy, one of the leading causes of miscarriages and birth defects in humans. Because the DSBs that serve as the initiating events of meiotic recombination pose a danger to genome integrity, the success of genome inheritance during meiosis requires cells to maintain a balance between the beneficial effects of COs and the potential harmful consequences of the process by which they are generated. Our goal is to understand the mechanisms that operate during meiosis to achieve this crucial balance. We are approaching this problem using the nematode C. elegans, a simple metazoan organism that is especially amenable to combining sophisticated cytological, genetic and genomic approaches in a single experimental system, and in which the events under study are particularly accessible and robust. The proposed work will exploit recent advances that provide the means to mark the sites of nascent CO events in live and fixed germ cells, to quantify the strength of CO interference, to manipulate the genome efficiently both at the DNA level and at the level of organismal ploidy, to visualize and reveal previously inaccessible organizational features of the DNA repair complexes assembled at recombination sites, and to obtain high-quality genome-wide information on chromatin organization from small numbers of cells. One goal is to elucidate the architecture and organization of DNA repair complexes at in vivo sites of meiotic recombination, with the objective of deducing and reconcilng relationships between the activities of individual recombination proteins/complexes and the in vivo functions of the collective CO recombination machinery. Another goal is to understand the mechanisms that promote and limit formation of meiotic COs, both at the level of designating recombination sites for a CO or non-CO fate and at the level of enforcing reliable execution of these fates. Finally, we will investigate processes that promote the formation of sufficient DSBs to guarantee CO formation and that efficiently channel these DSBs into homologous-recombination based repair pathways and away from error- prone mutagenic repair pathways.

项目概要：我们的长期目标是阐明基因重组如何有助于忠实遗传减数分裂期间的染色体，这是二倍体生物产生的特殊细胞分裂程序单倍体配子。减数分裂期间的染色体遗传依赖于双链DNA的形成断裂（DSB）和这些 DSB 子集的修复作为同源间交叉（CO）。未能形成 COs 导致染色体错误分离和随之而来的非整倍性，这是导致染色体异常的主要原因之一人类流产和出生缺陷。因为 DSB 作为减数分裂的起始事件重组对基因组完整性、减数分裂期间基因组遗传的成功构成危险需要细胞在CO的有益作用和潜在有害作用之间保持平衡它们产生的过程的后果。我们的目标是了解其中的机制在减数分裂期间进行操作以实现这一关键的平衡。我们正在使用以下方法来解决这个问题线虫 C. elegans，一种简单的后生动物生物体，特别适合结合复杂的在单一实验系统中进行细胞学、遗传学和基因组学方法，其中的事件正在研究的内容特别容易访问和强大。拟议的工作将利用最新的进展提供标记活体和固定生殖细胞中新生 CO 事件位点的方法，以量化 CO 干扰强度，在 DNA 水平和基因水平上有效地操纵基因组有机体倍性，可视化并揭示以前无法访问的 DNA 修复组织特征在重组位点组装复合物，并获得高质量的全基因组信息少量细胞的染色质组织。目标之一是阐明架构和在体内减数分裂重组位点组织 DNA 修复复合物，目的是推断和协调各个重组蛋白/复合物的活性之间的关系以及集体CO重组机制的体内功能。另一个目标是了解在指定重组水平上促进和限制减数分裂 CO 形成的机制 CO 或非 CO 命运的站点以及强制执行这些命运的可靠执行的级别。最后，我们将研究促进足够 DSB 形成以保证 CO 形成的过程，以及有效地将这些 DSB 引导至基于同源重组的修复途径并远离错误容易发生突变的修复途径。