小柯机器人

美国麻省理工学院Omar O. Abudayyeh、Fei Chen和Jonathan S. Gootenberg团队在研究中取得进展。他们使用RNA传感器ADAR进行可编程真核蛋白质合成。相关论文于2022年10月27日发表在《自然-生物技术》杂志上。

研究人员研发了一种可编程的RNA传感技术-可重新编程的ADAR传感器（RADARS），它利用作用于RNA腺苷脱氨酶（ADAR）的RNA编辑器通过内源性RNA转录物来门控转载蛋白的翻译。在转载蛋白编码基因上游的gudie中引入终止密码子，使翻译取决于内源性转录本与guide的结合，通过ADAR对终止密码子的编辑来进行翻译。通过系统的传感器工程，研究人员在传感器激活方面实现了277倍的改进，并针对各种货物蛋白（包括荧光素酶、荧光蛋白、重组酶和半胱天冬酶）设计了RADARS，从而能够在低于百万分之十三的转录水平对内源性转录本进行检测。

研究表明，RADARS可在表达的DNA或合成mRNA中发挥功能，并且具有外源性或内源性ADAR。RADARS可在多种情况下应用，包括跟踪转录状态、RNA感应诱导的细胞死亡、细胞类型鉴定以及对合成mRNA翻译的控制。

附：英文原文

Title: Programmable eukaryotic protein synthesis with RNA sensors by harnessing ADAR

Author: Jiang, Kaiyi, Koob, Jeremy, Chen, Xi Dawn, Krajeski, Rohan N., Zhang, Yifan, Volf, Verena, Zhou, Wenyuan, Sgrizzi, Samantha R., Villiger, Lukas, Gootenberg, Jonathan S., Chen, Fei, Abudayyeh, Omar O.

Issue&Volume: 2022-10-27

Abstract: Programmable approaches to sense and respond to the presence of specific RNAs in biological systems have broad applications in research, diagnostics, and therapeutics. Here we engineer a programmable RNA-sensing technology, reprogrammable ADAR sensors (RADARS), which harnesses RNA editing by adenosine deaminases acting on RNA (ADAR) to gate translation of a cargo protein by the presence of endogenous RNA transcripts. Introduction of a stop codon in a guide upstream of the cargo makes translation contingent on binding of an endogenous transcript to the guide, leading to ADAR editing of the stop codon and allowing translational readthrough. Through systematic sensor engineering, we achieve 277 fold improvement in sensor activation and engineer RADARS with diverse cargo proteins, including luciferases, fluorescent proteins, recombinases, and caspases, enabling detection sensitivity on endogenous transcripts expressed at levels as low as 13 transcripts per million. We show that RADARS are functional as either expressed DNA or synthetic mRNA and with either exogenous or endogenous ADAR. We apply RADARS in multiple contexts, including tracking transcriptional states, RNA-sensing-induced cell death, cell-type identification, and control of synthetic mRNA translation.

DOI: 10.1038/s41587-022-01534-5

Source: https://www.nature.com/articles/s41587-022-01534-5

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