Investigating the Roles of ADARs and A-to-I RNA Editing in Germline RNA Regulation

研究 ADAR 和 A-to-I RNA 编辑在种系 RNA 调控中的作用

• 批准号: 10705633
• 负责人: Emily Ann Erdmann
• 金额: $ 3.17万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-31 至 2026-08-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10705633
• 关键词: Address; Adenosine; Adult; Affect; Animals; Autoimmune Diseases; Binding; Biogenesis; Bioinformatics; Biology; CRISPR/Cas technology; Caenorhabditis elegans; Cell physiology; Complex; Coupled; Data; Deaminase; Deamination; Defect; Development; Dissection; Embryo; Embryonic Development; Ensure; Event; Gametogenesis; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Germ Cells; Goals; High-Throughput Nucleotide Sequencing; Human Pathology; Immunoprecipitation; Inosine; Investigation; Maintenance; Malignant Neoplasms; Messenger RNA; Metabolic Diseases; Methods; Molecular; Natural Immunity; Nematoda; Nerve Tissue; Nervous System; Northern Blotting; Pathway interactions; Phenotype; Physiological; Play; Population; Process; RNA; RNA Binding; RNA Editing; RNA-Binding Proteins; Regulation; Reproduction; Reproductive Biology; Research; Ribosomal RNA; Ribosomes; Role; Site; Small RNA; Stress; Surveys; Tissues; Training; Transcript; Translational Repression; Visualization; Work; differential expression; dsRNA adenosine deaminase; experience; experimental study; genome editing; high resolution imaging; in vivo imaging; insight; nervous system disorder; neural; offspring; programs; transcriptome sequencing; tumorigenesis

PROJECT SUMMARY The complex events of gametogenesis and early embryonic development require tight regulation to ensure proper reproduction and healthy offspring. To achieve this regulation while maintaining the plasticity necessary for embryogenesis, germ cells rely heavily on RNA-level regulation of gene expression. RNA regulation plays a number of important roles in germ cell development and maintenance, including maintenance of germ cell- specific gene programs, protection of the germline genome from foreign or damaging sequences, and translational repression and storage of maternal transcripts necessary for early embryonic development prior to the onset of zygotic transcription. A diverse network of RNA binding proteins and small RNA pathways govern this crucial RNA regulation, many of which have overlapping or redundant roles to ensure proper regulatory fine-tuning. Adenosine DeAminases that act on RNA (ADARs), are RNA binding proteins that can influence the cellular fate of transcripts either by binding to RNA or by catalyzing the deamination of Adenosine to Inosine, known as A-to-I RNA editing. ADARs are present in all animals and have been shown to play important roles in development, innate immunity, and oncogenesis. While the roles of ADARs in germ cells has not been explored, my own preliminary data suggests ADARs are highly expressed in the germline and can influence hundreds of germline transcripts. Additionally, previous work from other labs suggests that ADARs act along with small RNA pathways to perform functions necessary for proper reproduction. As such, I hypothesize that ADARs play a role in the extensive RNA regulation that takes place in the germline. My preliminary experiments investigating the effects of ADARs on germline transcripts suggest that ADARs play a role on regulating ribosome biogenesis. Additionally, I have found that many germline-edited transcripts are known maternal transcripts, which are translationally repressed throughout the germline and stored for loading into the embryo. For these reasons, I hypothesize that ADARs play a role in translational repression that occurs throughout the germline during gametogenesis. To address this hypothesis, I will expand upon my investigation of the effects of ADARs on germline transcripts as well as interactions with other germline RNA regulators (Aim 1). I will assess the impact of ADARs on germline ribosome biogenesis and translational activity (Aim 2). Finally, I will investigate the impact of ADAR editing on maternally loaded transcripts and early embryonic RNA populations (Aim 3). Through these experiments, I will gain an understanding of the effects of ADARs on germline transcripts, and a bigger-picture understanding of their role in germ cell development and maintenance, early embryonic development, and reproduction in general. This work will contribute to our understanding of the complex processes that facilitate successful and healthy reproduction.

项目摘要 配子发生和早期胚胎发育的复杂事件需要严格的调节以确保 适当的繁殖和健康的后代。为了实现此法规，同时保持必要的可塑性 对于胚胎发生，生殖细胞严重依赖基因表达的RNA级调节。 RNA调节播放 生殖细胞发育和维持中重要作用的数量，包括维持生殖细胞 特定的基因程序，保护种系基因组免受外国或破坏性序列的保护，以及 早期胚胎发育所需的母体成绩单的翻译压制和存储 合子转录的发作。 RNA结合蛋白和小的RNA途径的多种网络控制 这种关键的RNA调节，其中许多具有重叠或冗余的作用，以确保适当的调节 微调。作用于RNA（ADAR）的腺苷脱氨酶是RNA结合蛋白，可以影响 转录本的细胞命运是通过与RNA结合或通过催化腺苷对肌苷的脱氨 称为A到I RNA编辑。 ADAR都存在于所有动物中，并且已被证明在 发育，先天免疫和肿瘤发生。虽然ADAR在生殖细胞中的作用尚未 经过探索，我自己的初步数据表明，ADAR在种系中高度表达，可以影响 数百种种系成绩单。此外，其他实验室的先前工作表明ADAR的行动 具有小的RNA途径来执行适当繁殖所需的功能。因此，我假设 Adars在种系中发生的广泛的RNA法规中发挥了作用。我的初步 研究ADAR对种系成绩单的影响的实验表明，ADAR在 调节核糖体生物发生。此外，我发现许多种系编辑的成绩单是已知的 母体成绩单，在整个种系中被翻译抑制，并存储用于加载到 胚胎。由于这些原因，我假设ADAR在发生的翻译压抑中发挥了作用 在配子发生期间的整个种系。为了解决这一假设，我将扩大我的 研究ADAR对种系转录本的影响以及与其他种系RNA的相互作用 监管机构（目标1）。我将评估ADAR对种系核糖体生物发生和翻译的影响 活动（目标2）。最后，我将调查Adar编辑对母体负载的成绩单的影响 胚胎RNA种群（AIM 3）。通过这些实验，我将了解 关于种系成绩单的ADAR，以及对它们在生殖细胞发育中的作用的更大理解和 一般而言，维护，早期胚胎发育和繁殖。这项工作将有助于我们 了解促进成功繁殖的复杂过程。