Protein-PNA conjugates as conditional BRET platforms for life cell mRNA imaging and quantification

蛋白质-PNA 缀合物作为用于生命细胞 mRNA 成像和定量的条件 BRET 平台

• 批准号: 423428279
• 负责人: Dr. Frank Abendroth
• 金额: --
• 依托单位: Fachgebiet Chemische Biologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/423428279?language=en
• 关键词: Protein; PNA; conjugates; conditional; BRET

Within the framework of this proposal, a new highly sensitive imaging method will be developed to enable visualization and spatial quantification of low abundance mRNA isoforms in cells in real time. Despite the variety of existing RNA imaging methods, it is major hurdle to study mRNA isoforms in real time in living cells due to the lack of sensitivity of current strategies. Importantly, these low abundance transcripts formed mainly by alternative splicing contribute significantly to protein diversity and play a central role in development, neuronal plasticity, and nervous system disorders such as Alzheimer's, spinal muscular atrophy (SMA), etc. Based on the well-characterized mRNA isoforms responsible of SMA, we aim to develop a PNA-NanoBRET bioimaging platform, which allows follow gene expression in real time in living cells. This novel method will ultimately be tested in genetically modified cells and patient fibroblasts.

在该提议的框架内，将开发一种新的高灵敏度成像方法，以实现真实的时间内细胞中低丰度mRNA同种型的可视化和空间定量。尽管现有的RNA成像方法多种多样，但由于当前策略缺乏灵敏度，因此在活细胞中真实的时间内研究mRNA异构体是主要障碍。重要的是，这些主要通过选择性剪接形成的低丰度转录物对蛋白质多样性有显著贡献，并在发育、神经元可塑性和神经系统疾病如阿尔茨海默氏症、脊髓性肌萎缩症（SMA）等中发挥核心作用。基于负责SMA的良好表征的mRNA同种型，我们的目标是开发PNA-NanoBRET生物成像平台，其允许在活细胞中真实的时间内跟随基因表达。这种新方法最终将在转基因细胞和患者成纤维细胞中进行测试。