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

脊椎动物卵到胚胎转变过程中调节因子的鉴定

• 批准号: 10371345
• 负责人: Cara Moravec
• 金额: $ 9.1万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-12 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10371345
• 关键词: Actins; Address; Advisory Committees; Animals; Award; Candidate Disease Gene; Cell physiology; Cellular biology; Clinic; Complex; Congenital Abnormality; DNA; Defect; Deposition; Development; Embryo; Embryonic Development; Event; Female; Fertility; Gene Duplication; Genes; Genetic; Genome; Germ; Germ Cells; Goals; Human; Image; Impairment; Infertility; Mentors; Mentorship; Microtubules; Modeling; Molecular; Morphology; Mutation; Nature; Nuclear; Nuclear Envelope; Oocytes; Orthologous Gene; Pathway interactions; Pattern; Phase; Phenotype; Pregnancy; Process; RNA; Research; Research Personnel; Role; Spontaneous abortion; Techniques; Training; Vertebrates; Zebrafish; alpha Karyopherins; borealin; critical period; early pregnancy loss; egg; gene product; genetic approach; implantation; insight; mutant; novel; offspring; programs; skills; skills training; survivin; tool; whole genome

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.

项目摘要 在整个脊椎动物中，早期发育，在合子基因组激活之前，都取决于主要提供的 基因产品。这些产品将启动胚胎所必需的分子途径 开发编程。破坏这些产妇产品功能或稳定性的突变可能是 致命的胚胎致命。携带这些受损的孕妇基因的雌性不显示 明显的表型，但他们的后代将经历与其遗传学无关的异常发展。 尽管母体效应基因在早期发育中的重要性，但我们仅确定了 通过遗传方法的一小部分母体效应基因。在人类中，母亲表达的缺陷 预计基因会导致植入或早期妊娠丧失失败。梅奥诊所估计十个 到20％的已知妊娠被流产，但是这一百分比可能更高，因为 早期流产可能不会被发现。该建议旨在解决主要表达基因的作用 在鸡蛋过渡期间。具体而言，在AIM 1（K99）中，我将执行实时的亚细胞成像 表征主要表达的染色体乘客复合物在调节动态方面的作用 微管依赖性种质聚集的性质。在AIM 2（K99）中，我将探索一个 在调节核包膜中，母体表达的Importin-α，kpna7在卵到胚胎期间重新组装 过渡。在我的r00年中，我将使用大型的母室屏幕来识别新颖的调节器 鸡蛋到embryo的过渡。初步研究表明，这是确定的有效方法 主要表达基因在发育中的作用。通过将这个较清晰的屏幕与现场直播相结合 早期开发所需的动态过程的亚细胞成像，我将开始理解 新确定的生物作用基因在早期胚胎中的作用。我的K99培训将受到特殊的指导 导师，弗朗西斯科·贝利格里（Francisco Pelegri）博士和威廉·伯门（William Bement）博士，以及咨询委员会，所有这些都将提供 支持和心态使我能够过渡到独立性。我已建立的通用的结合 与Bement博士一起编辑技能和实时亚细胞成像的培训将为我提供独特的技能。 需要研究主要表达基因在卵到胚胎过渡中的作用。 K99/R00 奖项将使我能够成为孕产妇在开发过程中角色的独立研究者 并将提供对不孕症和早期妊娠丧失的遗传学的理解。