Modular, programmable RNA sensing using ADAR editing in living cells
Modular, programmable RNA sensing using ADAR editing in living cells
复制标题
DOI：
10.1038/s41587-022-01493-x
复制
发表时间：
2022-10-05
影响因子：
46.9
通讯作者：
Gao, Xiaojing J.
中科院分区：
文献类型：
--
作者：
Kaseniit, K. Eerik;Katz, Noa;Gao, Xiaojing J.
通过利用内源性ADAR酶在活细胞中检测到特定的RNA。随着单细胞转录本的日益可用，RNA签名为靶向活细胞提供了一个有希望的基础。分子RNA传感器将能够在不同的环境中研究特定细胞类型/状态并对其进行治疗干预，特别是在人类患者和非模式生物中。在这里，我们描述了一个模块化的，可编程的活体RNA传感系统，使用作用于RNA的腺苷脱氨酶(雷达)。我们验证并扩展我们的基本设计，表征其性能，并分析其与人类和小鼠转录本的兼容性。我们确定了提高产出水平和改善动态范围的战略。此外，我们还展示了雷达能够实现紧凑和逻辑。除了对转录水平做出反应外，雷达还可以区分与疾病相关的转录身份序列变化，如点突变和融合。最后，我们证明了雷达是一个自给自足的系统，有可能在不同的生物体中发挥作用。
Specific RNAs are detected in live cells by harnessing endogenous ADAR enzymes.With the increasing availability of single-cell transcriptomes, RNA signatures offer a promising basis for targeting living cells. Molecular RNA sensors would enable the study of and therapeutic interventions for specific cell types/states in diverse contexts, particularly in human patients and non-model organisms. Here we describe a modular, programmable system for live RNA sensing using adenosine deaminases acting on RNA (RADAR). We validate, and then expand, our basic design, characterize its performance, and analyze its compatibility with human and mouse transcriptomes. We identify strategies to boost output levels and improve the dynamic range. Additionally, we show that RADAR enables compact AND logic. In addition to responding to transcript levels, RADAR can distinguish disease-relevant sequence alterations of transcript identities, such as point mutations and fusions. Finally, we demonstrate that RADAR is a self-contained system with the potential to function in diverse organisms.