利用工程化的光控 RNA 结合蛋白对 RNA 功能和代谢进行光遗传学控制。
Optogenetic control of RNA function and metabolism using engineered light-switchable RNA-binding proteins.
机构信息
Optogenetics & Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
School of Pharmacy, East China University of Science and Technology, Shanghai, China.
出版信息
Nat Biotechnol. 2022 May;40(5):779-786. doi: 10.1038/s41587-021-01112-1. Epub 2022 Jan 3.
RNA-binding proteins (RBPs) play an essential role in regulating the function of RNAs in a cellular context, but our ability to control RBP activity in time and space is limited. Here, we describe the engineering of LicV, a photoswitchable RBP that binds to a specific RNA sequence in response to blue light irradiation. When fused to various RNA effectors, LicV allows for optogenetic control of RNA localization, splicing, translation and stability in cell culture. Furthermore, LicV-assisted CRISPR-Cas systems allow for efficient and tunable photoswitchable regulation of transcription and genomic locus labeling. These data demonstrate that the photoswitchable RBP LicV can serve as a programmable scaffold for the spatiotemporal control of synthetic RNA effectors.
RNA 结合蛋白（RBPs）在调节细胞环境中 RNA 的功能方面发挥着重要作用，但我们控制 RBP 活性的时间和空间的能力是有限的。在这里，我们描述了 LicV 的工程设计，它是一种光致变色的 RBP，可在蓝光照射下响应特定的 RNA 序列。当与各种 RNA 效应子融合时，LicV 可实现细胞培养中 RNA 定位、剪接、翻译和稳定性的光遗传学控制。此外，LicV 辅助的 CRISPR-Cas 系统可实现转录和基因组位点标记的高效且可调的光开关调节。这些数据表明，光致变色 RBP LicV 可用作合成 RNA 效应子的时空控制可编程支架。
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