Cell Cycle Regulation In C. elegans

线虫的细胞周期调控

基本信息

批准号：
8553386
负责人：
Andy Golden
金额：
$ 14.82万
依托单位：
NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

项目摘要

Our lab is interested in the process of eggshell development and its influence on early embryonic development. It has been observed that defects in eggshell development always result in embryonic lethality. It thus remains unclear whether the eggshell provides solely a structural support for the developing embryo or if it also plays a signaling role necessary for early development. It is known that proteins important for eggshell synthesis are carefully regulated and degraded during early development. In an attempt to better understand the role of the eggshell, its synthesis, and what maternal and paternal factors influence its function, we are studying a number of embryonic lethal mutants that have been reported to have eggshell defects. Genetic screens carried out in the early 1980s identified a large collection of interesting mutants, the majority of which were not further studied nor molecularly cloned. With the advent of whole genome sequencing, it is now possible in a very short time (and for very little research funds) to molecularly identify the mutations responsible for these interesting phenotypes. We are currently characterizing all 20 of these mutants in greater detail. For the whole genome sequencing, we are first out-crossing the strains one time to an Hawaiian C. elegans strain and re-isolating our embryonic lethal mutant. Those animals are then subjected to whole genome sequencing. The genomic regions free of Hawaiian SNPs are then identified as the regions bearing our mutation of interest. We then will RNAi candidate genes in these regions to identify the responsible gene. Those mutants with interesting phenotypes will be further characterized. It also remains possible that we may purse specific mutants because of the gene that is disrupted, with a specific focus on genes with human orthologs.

我们的实验室对蛋壳发育的过程及其对早期胚胎发育的影响感兴趣。已经观察到蛋壳发育中的缺陷总是导致胚胎致死性。因此，尚不清楚蛋壳是否仅提供对发育中的胚胎的结构支持，还是还起着早期发育所需的信号传导作用。众所周知，在早期发育过程中，对蛋壳合成重要的蛋白质仔细调节和降解。为了更好地了解蛋壳的作用，其合成以及母亲和父亲因素影响其功能，我们正在研究许多据报道具有蛋壳缺陷的胚胎致死突变体。在1980年代初进行的遗传筛选确定了大量有趣的突变体，其中大多数没有得到进一步研究或分子克隆。随着整个基因组测序的出现，现在可以在很短的时间内（很少的研究基金）来识别负责这些有趣表型的突变。我们目前正在更详细地描述所有这些突变体中的所有20个突变体。对于整个基因组测序，我们首先一次将菌株越过夏威夷秀丽隐杆线虫菌株并重新分配我们的胚胎致死突变体。然后，将这些动物进行整个基因组测序。然后将不含夏威夷SNP的基因组区域确定为带有我们感兴趣突变的区域。然后，我们将在这些区域中RNAi候选基因识别负责的基因。那些具有有趣表型的突变体将进一步表征。由于被破坏的基因，我们也可能会推动特定的突变体，并特别关注具有人类直系同源物的基因。