Artificial dynamic structure ensemble- guided rational design of a universal RNA aptamer-based sensing tag

• 成果类型: Article
• 署名作者: Hou, Jianing; Guo, Pei; Wang, Junyan; Han, Da; Tan, Weihong
• 署名单位: Shanghai Jiao Tong University; Chinese Academy of Sciences; Hangzhou Institute of Medicine, CAS
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-12749
• DOI: 10.1073/pnas.2414793121
• 发表日期: 2024-12-24
• 关键词: fluorescence; parameters; knockout; protein; motifs

摘要

Artificially functional RNAs, such as fluorogenic RNA aptamer (FRApt)-based biosensing tag, represent significant advancements in various biological applications but are limited by the lack of insight into dynamic structure ensembles and universal design concepts. Through the development of an artificial RNA structure ensemble, we rationally established an RNA reconstitution model, SSPepper- Apt, to generate a universal fluorogenic RNA biosensing tag. By utilizing various target- recognizing RNA motifs, SSPepper-Apt enables the modular generation of sensing tags for low- background, highly selective imaging of metabolites, peptides, and proteins in living cells. Additionally, by employing single guide RNA (sgRNA) as the recognition RNA motif, SSPepper-Apt generates fluorescence in both CRISPR- mediated imaging and gene editing only when the Cas9-sgRNA complex is successfully assembled; therefore, it can be an effective sgRNA screening tool for gene editing. Our fluorogenic RNA- sensing tag provides a universal approach for constructing functional RNA systems, avoiding the laborious and time- consuming process of sequence combination, and expanding the application of synthetic biological tools.