RUI: Studying B-bodies, a novel type of nuclear domains: from organization to practical exploitation.

RUI：研究 B 体，一种新型核域：从组织到实际利用。

• 批准号: 2042814
• 负责人: Anton Bryantsev
• 金额: $ 41.98万
• 依托单位: Kennesaw State University Research and Service Foundation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2042814&HistoricalAwards=false
• 关键词: RUI; Studying; bodies; novel; type

This project studies basic principles that govern organization of the cell nucleus. The nucleus has a special importance for biotechnology because it stores genetic information that controls traits of cells and organisms. Continuous success of biotechnology greatly depends on better understanding of the processes inside of the nucleus. This project will specifically focus on studying the sorting mechanism that enables certain proteins to form distinct and isolated structures within the nucleus. In a separate set of experiments, the information learned in this project will be applied to improve the current biotechnology methods. The desired outcome is to make genetic manipulations more efficient and safer. As a broader impact, this project will offer an educational platform at Kennesaw State University (KSU) to train undergraduate and graduate students with contemporary methods of molecular biology and genetic engineering. KSU is a dynamically developing, predominantly undergraduate institution that strives to enhance its research environment and expand graduate programs.In the absence of internal membranes in the nucleus, nuclear proteins are organized into compact, distinct areas with sharp boundaries that are called nuclear domains (NDs). The regulation and functional significance of protein sorting into NDs is not clear. This project will use a newly discovered Drosophila ND called a B-body and its resident RNA-binding protein called Bruno, as a model to study ND assembly and function. It is hypothesized that a combination of Bruno’s protein sequence and non-coding RNA is sufficient to create B-bodies. Molecular biology techniques, including mutagenesis and in situ RNA hybridization, will be applied to identify the essential components to reconstitute B-bodies. Next, B-bodies will be tested for their role as safe overflow sites for ectopically expressed nuclear proteins. Heterologous proteins including DNA enzymes and transcription factors will be targeted to B-bodies. Biochemical assays and genetic reporters will quantitatively indicate whether protein trafficking into B-bodies can reduce non-specific effects and toxicity that often plague overexpression experiments.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这个项目研究支配细胞核组织的基本原理。细胞核对生物技术具有特殊的重要性，因为它储存了控制细胞和生物体性状的遗传信息。 生物技术的持续成功在很大程度上取决于对细胞核内部过程的更好理解。 该项目将特别关注研究使某些蛋白质能够在细胞核内形成独特和孤立结构的分选机制。 在一组单独的实验中，在这个项目中学到的信息将被应用于改善目前的生物技术方法。 期望的结果是使基因操作更有效和更安全。 作为一个更广泛的影响，该项目将提供一个教育平台，在肯尼索州立大学（KSU）培养本科生和研究生与现代方法的分子生物学和遗传工程。 KSU是一个充满活力的发展，主要是本科院校，努力提高其研究环境和扩大研究生课程。在细胞核中没有内膜的情况下，核蛋白被组织成紧凑，清晰的区域，具有清晰的边界，称为核域（ND）。 蛋白分选成ND的调控和功能意义尚不清楚。该项目将使用一种新发现的果蝇ND（称为B体）及其驻留的RNA结合蛋白（称为Bruno）作为研究ND组装和功能的模型。 据推测，布鲁诺的蛋白质序列和非编码RNA的组合足以产生B体。 分子生物学技术，包括诱变和原位RNA杂交，将用于鉴定重组B体的必要组分。 接下来，将测试B体作为异位表达核蛋白的安全溢出位点的作用。 包括DNA酶和转录因子的杂合蛋白将靶向B体。 生化分析和基因报告将定量地表明蛋白质运输到B体是否可以减少非特异性的影响和毒性，往往困扰过表达experiments.This奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。