Arabidopsis 2010: Saturating the Arabidopsis Knockout Collection with Lethals

拟南芥研究界的一个目标是确定每个注释基因的敲除。目前收集的纯合子敲除突变体中缺失的是由于配子体致死性导致突变种子流产或无法形成的关键基因。这个项目的目的是协调各方的努力，以填补这一空白。该结果将为未来的实验提供宝贵的资源，并为破坏拟南芥中非冗余必需基因的后果提供基准信息。主要研究人员已经建立了一个强大的数据库（www.seedgenes.org），其中包含拟南芥种子发育所需的300个EMB基因。计划采用三种方法来扩大这一社区资源。第一种涉及基因，现有的敲除项目发现插入系具有确认的杂合子，但没有纯合子敲除。将根据已知的序列冗余、同源性、表达模式以及与已知必需基因的共同特征，对该列表进行筛选，以确定有希望成为胚胎或配子体致死基因的候选基因。然后将选定的品系进行种植、基因分型和确认。他们将筛选在种子和配子体发育缺陷。目标是在三年内分析450到600个有希望的插入线。第二种方法涉及收集在其他实验室发现的致命突变体的信息。这种方法在种子突变体中效果很好，并将在未来扩展到包括假定的配子体突变体。第三种方法涉及在未发现或无法确认插入的剩余基因中识别有希望的必要基因候选者。这种方法将产生雄性和雌性配子体发育所需的候选基因列表。这些联合方法的结果将被纳入SeedGenes和TAIR数据库。在其他模式生物中具有未知功能和没有同源物但具有确定的种子表型的基因将进行最详细的检查。这些努力将通过关注已知的关键基因，增强我们对拟南芥未知基因的理解。该项目更广泛的教育影响包括高中生参与拟南芥突变体和植物分子遗传学的研究活动和推广计划，涉及地区高中，斯蒂尔沃特初中的扩展研究计划，以及俄克拉何马州科学与数学学院与俄勒冈州立大学植物生物技术网络合作。这些项目在上一个赠款期间成功启动。

One goal of the Arabidopsis research community is to identify a knockout for every annotated gene. Missing from the current collection of homozygous knockout mutants are the essential genes for which mutant seeds abort or fail to form because of gametophytic lethality. The aim of this project is to coordinate efforts to fill this gap in the knockout collection. The result will be a valuable resource for future experiments and benchmark information on the consequences of disrupting non-redundant, essential genes in Arabidopsis. The principal investigators have already established a robust database (www.seedgenes.org) of 300 EMB genes required for seed development in Arabidopsis. Three approaches are planned to expand upon this community resource. The first involves genes where the existing knock-out project finds insertion lines with confirmed heterozygotes but no homozygous knockouts. This list will be filtered to identify genes that represent promising candidates for embryo or gametophytic lethals based on known sequence redundancy, orthology, expression patterns, and shared features with known essential genes. Selected lines will then be planted, genotyped, and confirmed. They will be screened for defects in seed and gametophyte development. The goal is to analyze 450 to 600 promising insertion lines over three years. The second approach involves collecting information on lethal mutants identified in other laboratories. This approach has worked well for seed mutants and will expand in the future to include putative gametophytic mutants. The third approach involves identification of promising candidates for essential genes among those remaining genes for which insertions have not been found or cannot be confirmed. This approach will generate a list of candidate genes that are required for both male and female gametophyte development. Results of these combined approaches will be incorporated into the SeedGenes and TAIR databases. Genes with unknown functions and without orthologs in other model organisms but with a confirmed seed phenotype will be examined in most detail. These efforts will enhance our understanding of the unknowns in Arabidopsis by focusing on genes known to be essential. The broader educational impact of this project includes participation of high school students in research activities and outreach programs on Arabidopsis mutants and plant molecular genetics involving regional high schools, the Extended Studies Program at Stillwater Junior High School, and the Oklahoma School for Science and Mathematics in conjunction with the OSU Plant Biotechnology Network. These programs were successfully initiated during the previous grant period.