Collaborative Research: Ideas Lab: Rational Design of Noncoding RNA for Epigenetic Signal Amplification

合作研究：创意实验室：用于表观遗传信号放大的非编码 RNA 的合理设计

• 批准号: 2243666
• 负责人: Keriayn Smith
• 金额: $ 66.58万
• 依托单位: University of North Carolina at Chapel Hill
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-15 至 2027-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2243666&HistoricalAwards=false
• 关键词: Collaborative; Research; Ideas; Lab; Rational

Considerable progress has been made to illuminate the dark matter of the genome - those segments that do not encode for protein. Some of these noncoding regions are transcribed to produce long noncoding RNAs (lncRNAs) that are processed similarly to protein-encoding messenger RNAs. Although they are often found only at low amounts within a cell, lncRNAs have been implicated in a myriad of biological processes including cell growth and survival, cell identity and environmental interactions, as well as numerous human and animal diseases. The next frontier will be to tap into the biotechnology potentially inherent to these lncRNAs. This project will use a synthetic, ‘understanding by building’ approach to rationally design and use lncRNAs to control large swaths of chromosomal DNA. Broad utility will be demonstrated by using this new RNA technology in human cells and in yeast (fungi). Engineering lncRNAs to control their activity will not only contribute to fundamental understanding of these enigmatic RNAs, but also tap the vast potential to generate useful biomolecules in agricultural and medical applications across species. The project includes uniquely tailored programming to support STEM workforce development for participants ranging from high school students to postdoctoral fellows. New virtual and hands-on activities focused on RNA biology will be integrated with successful STEM outreach programs to ensure strong impact.This project seeks to define the minimal features that are required for large-scale silencing mediated by lncRNA, building off of the well-characterized Xic locus and its associated lncRNA, Xist, which is responsible for silencing one of the X chromosomes in female mammals and marsupials. The ultimate goal is to gain fundamental understanding of the mechanisms by which lncRNAs can exert silencing effects at a chromosomal scale despite being expressed at a low level in the cell. To tackle this, the investigators will first test the effects of different repeats on silencing ability in a mammalian cell culture system. They will then attempt to silence autosomal loci by integrating synthetic lncRNAs into other chromosomes, or via CRISPR-based targeting. We will use these data to parameterize mathematical models to better understand the dynamics and limitations of the silencing process. They will also create a stand-alone silencing system in haploid yeast cells using components of the Xist system. This will enable detailed genetic manipulations to further test and optimize the silencing system using the power of yeast genetics. Finally, the project will use the data to guide rational design of synthetic silencing systems that can be readily deployed in mammalian cells to achieve targeted and tunable silencing. Overall, the investigators expect to push the boundaries of the current understanding of lncRNA functions in the nucleus, with anticipated outcomes to include defining the minimal components and lncRNA levels required for stable and specific epigenetic regulation. By reconstituting functional components from the ground-up, they expect to validate, challenge, and/or expand fundamental mechanics involving RNAs. Given that tunable lncRNA modulation could be a broadly applicable biotechnology, this work has exciting implications for bioengineering.This collaborative project resulted from an NSF-sponsored Ideas Lab. It will be co-funded by the Emerging Frontiers Program and the Division of Molecular and Cellular Biosciences.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.

基因组的深色物质已被转录，以产生长期的非编码RNA（LNCRNA），这些RNA被处理到蛋白质编码的Messenger RNA中。随着许多人类和动物疾病的范围。人类的LS技术（真菌）。该项目旨在定义Lncrna大规模沉默所必需的微型S，在良好的XIC基因座及其相关的LNCRNA上建立，这是为了使女性哺乳动物和象征的X染色体之一沉默最终目标是通过LNCRNA获得对机制的基本理解，可以在NVestigators上发挥作用，首先要测试扩散能力的差异对沉默能力的差异，然后试图通过Integrat Synthetic LNCRNAS沉默的常染色体lncrnas对其他染色体进行沉默。沉默过程的动力和局限性。可调节的lncRNA在核中的当前理解，以最小的成分和特定的表观遗传学调节定义了通过地面的稳定和特定的表观遗传组件此协作计划和R和Cellular Biosciences的部门。该奖项反映了NSF'SFLEF的使命，并通过使用脚趾脚趾的脚趾脚趾的脚趾进行评估来获得不值得的支持和更广泛的影响审查标准。