Identification of regulators in the vertebrate egg-to-embryo transition

项目摘要 在脊椎动物中，合子基因组激活之前的早期发育依赖于母体供应 基因产品。这些产物将启动胚胎所必需的分子途径。 发展规划。破坏这些母体产物的功能或稳定性的突变可能是 对发育中的胚胎是致命的。携带这些受损的母性效应基因的女性不会表现出 明显的表型，但他们的后代将经历独立于他们的遗传的异常发育。 尽管母性效应基因在早期发育中很重要，但我们只确定了 通过遗传方法获得一小部分母性效应基因。在人类中，母性表达的缺陷 基因预计会导致着床失败或早孕丢失。梅奥诊所估计有10个 到20%的已知怀孕是流产的，但这个比例可能要高得多，因为 早期流产可能不会被发现。这项提案旨在解决母体表达的基因的作用 在卵子到胚胎的转变过程中。具体地说，在目标1(K99)中，我将执行实时亚细胞成像以 描述母体表达的染色体乘客复合体在调节动态过程中的作用 依赖微管的种质聚集的性质。在目标2(K99)中，我将探索 Importin-α、KpNA7在母体表达调控卵子到胚胎核膜重组中的作用 过渡。在我的R00年里，我将使用一个大尺寸的母性清晰的屏幕来识别新的调节器 从卵子到胚胎的转变。初步研究表明，这是一种有效的确定 母体表达基因在发育中的作用。通过将这款母性清爽的屏幕与现场直播相结合 亚细胞成像是早期发育所需的动态过程，我将开始了解 新发现的母体效应基因在早期胚胎中的作用。我的K99培训将由卓越的 导师，弗朗西斯科·佩雷格里博士和威廉·贝门特博士，以及一个咨询委员会，他们都将提供 支持和指导使我能够过渡到独立。我已有的基因组合 编辑技能和与Bement博士一起接受的实时亚细胞成像培训将使我获得独特的技能集，即 需要研究母体表达的基因在卵子到胚胎转化中的作用。K99/R00 获奖将使我成为一名独立的调查者，研究母体产品在发育过程中的作用 并将提供对不孕症和早孕丢失的遗传学的理解。

Project Summary Across vertebrates, early development, prior to zygotic genome activation, is dependent on maternally supplied gene products. These products will initiate molecular pathways that are necessary for the embryonic developmental programing. Mutations that disrupt the function or stability of these maternal products can be lethal to the developing embryo. Females who carry these impaired maternal-effect genes do not display an overt phenotype, but their offspring will undergo abnormal development that is independent of their genetics. Despite the importance of maternal-effect genes in early development, we have only determined the function of a small subset of maternal-effect genes via genetic approaches. In humans, defects in maternally expressed genes are expected to result in failed implantation or early pregnancy loss. The Mayo Clinic estimates that ten to twenty percent of known pregnancies are miscarried, but this percentage is likely significantly higher because early miscarriages can go undetected. This proposal aims to address the role of maternally expressed genes during the egg to embryo transition. Specifically, in Aim 1 (K99), I will perform live subcellular imaging to characterize the role of a maternally expressed chromosomal passenger complex in regulating the dynamic nature of microtubule-dependent germplasm aggregation. In Aim 2 (K99), I will be exploring the role of a maternally expressed Importin-α, Kpna7 in regulating nuclear envelope reassembly during the egg-to-embryo transition. During my R00 years, I will use a large scale maternal-crispant screen to identify novel regulators of the egg-to-embryo transition. Preliminary studies have suggested that this is an efficient way to determine the role of maternally expressed genes in development. By combining this maternal crispant screen with live subcellular imaging of the dynamic processes that are required for early development, I will start understanding the role of newly identified maternal effect genes in early embryo. My K99 training will be guided by exceptional mentors, Dr. Francisco Pelegri and Dr. William Bement, and an advisory committee, all of whom will provide support and mentorship allowing me to transition into independence. The combination of my established genetic editing skill and the training in live subcellular imaging with Dr. Bement will give me the unique skill set that is required to research the role of the maternally expressed genes in the egg-to-embryo transition. The K99/R00 award will allow me to become an independent investigator in the role of maternal products during development and will provide understanding into the genetics of infertility and early pregnancy loss.