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: 10663791
• 负责人: Cara Moravec
• 金额: $ 8.43万
• 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/10663791
• 关键词: Actins; Address; Advisory Committees; Animals; Award; Candidate Disease Gene; Cell physiology; Cellular biology; Clinic; Complex; Congenital Abnormality; DNA; Defect; Deposition; Development; Dissociation; Embryo; Embryonic Development; Event; Female; Fertility; Gene Duplication; Generations; 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; segregation; skills; 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.

项目概要 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 这些产物将启动胚胎必需的分子途径。 破坏这些母体产物的功能或稳定性的突变可能是。 携带这些受损母体效应基因的雌性不会表现出对胚胎发育的致命性。 明显的表型，但他们的后代将经历与其遗传无关的异常发育。 尽管母体效应基因在早期发育中很重要，但我们只确定了母体效应基因的功能 通过遗传方法产生的一小部分母体效应基因，在人类中，母体表达的缺陷。 梅奥诊所估计有十种基因会导致植入失败或早期流产。 已知怀孕的百分之二十是流产的，但这个百分比可能要高得多，因为 早期流产可能未被发现。该提案旨在解决母体表达基因的作用。 具体来说，在目标 1 (K99) 中，我将进行活体亚细胞成像。 描述母体表达的染色体乘客复合物在调节动态中的作用 在目标 2 (K99) 中，我将探索微管依赖性种质聚集的作用。 母体表达的 Importin-α、Kpna7 在卵子到胚胎过程中调节核膜重组 在我的 R00 岁月里，我将使用大规模的母体脆皮筛选来识别新的调节因子。 初步研究表明，这是确定卵子到胚胎的有效方法。 通过将母体脆性筛选与活体结合，研究母体表达基因在发育中的作用。 早期发育所需的动态过程的亚细胞成像，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将受到特殊的指导。 导师 Francisco Pelegri 博士和 William Bement 博士以及咨询委员会，他们都将提供 支持和指导让我能够过渡到独立。 编辑技能以及 Bement 博士的活体亚细胞成像培训将为我提供独特的技能， 研究母体表达基因在卵子到胚胎转变中的作用所需。 该奖项将使我能够成为一名独立调查员，研究母体产品在开发过程中的作用 并将提供对不孕症和早期妊娠流产的遗传学的了解。