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Genetically encoded RNA-based sensors with Pepper fluorogenic aptamer
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DOI：10.1093/nar/gkad620.png)
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摘要
En 中文
Sensors to measure the abundance and signaling of intracellular molecules are crucial for understanding their physiological functions. Although conventional fluorescent protein-based sensors have been designed, RNA-based sensors are promising imaging tools. Numerous RNA-based sensors have been developed. These sensors typically contain RNA G-quadruplex (RG4) motifs and thus may be suboptimal in living cells. Here we describe RNA-based sensors based on Pepper, a fluorogenic RNA without an RG4 motif. With Pepper, we engineered various sensors for metabolites, synthetic compounds, proteins and metal ions in vitro and in living cells. In addition, these sensors show high activation and selectivity, demonstrating their universality and robustness. In the case of sensors responding to S-adenosylmethionine (SAM), a metabolite produced by methionine adenosyltransferase (MATase), we showed that our sensors exhibited positively correlated fluorescence responding to different SAM levels. Importantly, we revealed the SAM biosynthesis pathway and monitored MATase activity and gene expression spatiotemporally in living individual human cells. Additionally, we constructed a ratiometric SAM sensor to determine the inhibition efficacy of a MATase inhibitor in living cells. Together, these sensors comprising Pepper provide a useful platform for imaging diverse cellular targets and their signaling pathway.
Keyword:
GREEN FLUORESCENT PROTEIN
CRYSTAL-STRUCTURE
GENE-EXPRESSION
LIVING CELLS
TRANSLATION
METABOLITES
NANODEVICES
SELECTION
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期刊
IF:
13.1
论文数:
3.6W
被引数:
29.0W
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Imaging Intracellular S-Adenosyl Methionine Dynamics in Live Mammalian Cells with a Genetically Encoded Red Fluorescent RNA-Based Sensor使用基于遗传编码的红色荧光RNA的传感器对活哺乳动物细胞中的细胞内S-腺苷甲硫氨酸动力学进行成像OAAI
OAAI分享收藏Genetically Encoded Ratiometric RNA-Based Sensors for Quantitative Imaging of Small Molecules in Living Cells基于遗传编码的比率RNA传感器，用于活细胞中小分子的定量成像OAAI
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