Genetic Dissection of Germ Cell Differentiation and Function

生殖细胞分化和功能的基因剖析

• 批准号: 10330396
• 负责人: Michael Buszczak
• 金额: $ 26.58万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10330396
• 关键词: Adopted; Area; Back; Biogenesis; CRISPR/Cas technology; Cell Differentiation process; Cell physiology; Cells; Cellular biology; DNA Repair; DNA Transposable Elements; Development; Dissection; Drosophila genome; Drosophila genus; Embryonic Development; Exhibits; Foundations; Genes; Genetic; Genetic Translation; Genome; Genome Stability; Germ Cells; Germ cell tumor; Gonadal structure; Hand; Haploidy; Health; Human; Medical; Meiosis; Methods; Modeling; Molecular; Mutate; Mutation; Nuclear; Ovary; Pathway interactions; Peptide Hydrolases; Phenotype; Process; Regulation; Ribosomes; Science; Small RNA; Solid; Totipotency; Work; base; epigenome; fascinate; innovation; insight; screening; tool

Summary The study of germ cells has shaped our understanding of many basic fundamental processes across different species. Germ cells share a number of features that have long fascinated biologists. These cells undergo meiosis to form haploid gametes, they are exceptionally good at repairing DNA damage, they utilize a number of small RNA pathways to silence transposable elements, and they reprogram their epigenome back to a state that supports totipotency. Here, we proposed to use the Drosophila ovary as a model to continue to gain insights into genome stability, germ cell differentiation, and the cell-specific regulation of mRNA translation and ribosome biogenesis/turnover. Over the last five years, we have adopted and optimized a number of innovative CRISPR-Cas9- and recombineering-based methods for manipulating the Drosophila genome. Using these approaches, we have mutated and/or tagged over 100 genes that exhibit enriched expression in Drosophila gonads. This work provides a solid foundation for our planned efforts over the next five years. We will focus on a number of different but related areas. We will continue to characterize the highly conserved Germ Cell Nuclear Acidic Peptidase (GCNA) gene and its function in protecting the integrity of germ cells across species. We will also continue to characterize how cytoplasmic Rbfox1 controls early germ cell development. Our previous screening efforts have identified a small number of mutations that exhibit germ cell tumor formation or germ cell loss phenotypes. The molecule function of the disrupted genes will be characterized using the tools and methods we have in hand. Lastly, we are in the process of generating a number of innovative tools that will allow us to better assess ribosome biogenesis and turnover during germ cell development and early embryogenesis. We are very excited by this proposed work and believe the successful completion of these projects will have a positive impact on our understanding of germ cell biology and other molecular processes that impact human health.

概括 生殖细胞的研究塑造了我们对跨越许多基本基本过程的理解 不同的物种。生殖细胞具有许多长期吸引生物学家的特征。这些细胞 经历减数分裂以形成单倍配子，它们非常擅长修复DNA损伤，它们 利用许多小的RNA途径来沉默转座元素，然后重新编程 表观基因组回到支持能力的状态。在这里，我们建议将果蝇卵巢作为 一个模型继续深入了解基因组稳定性，生殖细胞分化和细胞特异性 MRNA翻译和核糖体生物发生/更新的调节。在过去的五年中，我们有 采用并优化了许多创新的CRISPR-CAS9和基于重新组合的方法 操纵果蝇基因组。使用这些方法，我们已经突变和/或标记了100多个 在果蝇性腺中表现出富集表达的基因。这项工作为 我们计划在未来五年内进行的努力。我们将专注于许多不同但相关的领域。我们 将继续表征高度保守的生殖细胞核酸肽酶（GCNA）基因和 它在保护跨物种的生殖细胞完整性方面的功能。我们还将继续描述 细胞质RBFOX1如何控制早期生殖细胞发育。我们以前的筛选工作 鉴定出少量表现出生殖细胞肿瘤或生殖细胞损失的突变 表型。破坏基因的分子函数将使用工具和 我们手头的方法。最后，我们正在生成许多创新工具 将使我们能够更好地评估生殖细胞发育期间和早期的核糖体生物发生和周转 胚胎发生。我们对这项拟议的工作感到非常兴奋，并相信成功完成 这些项目将对我们对生殖细胞生物学和其他分子的理解产生积极影响 影响人类健康的过程。