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 基因产物。这些产品将启动胚胎发育所需的分子途径 发展性规划。破坏这些母体产物的功能或稳定性的突变可能是 对发育中的胚胎具有致命性。携带这些受损母体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 通过遗传方法获得的一小部分母体效应基因。在人类中，母性表达缺陷 基因预计会导致植入失败或早期妊娠流产。梅奥诊所估计，十 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 在卵子到胚胎的转变过程中。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 微管依赖性种质聚集的性质。在目标 2 (K99) 中，我将探索 母体表达的 Importin-α、Kpna7 在卵子到胚胎过程中调节核膜重组 过渡。在我的 R00 岁月里，我将使用大型母体脆皮屏幕来识别新的调节因子 卵子到胚胎的转变。初步研究表明，这是确定 母系表达基因在发育中的作用。通过将这个产妇脆屏与现场直播相结合 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。我的 K99 培训将由卓越的指导 导师 Francisco Pelegri 博士和 William Bement 博士以及咨询委员会，他们都将提供 支持和指导使我能够过渡到独立。我已建立的遗传基因的组合 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， 需要研究母体表达基因在卵子到胚胎转变中的作用。 K99/R00 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。