Genetically Encoded Dual-Color Light-Up RNA Sensor Enabled Ratiometric Imaging of MicroRNA

MicroRNAs (miRNAs) play essential roles in regulating gene expression and cell fate. However, it remains a great challenge to image miRNAs with high accuracy in living cells. Here, we report a novel genetically encoded dual-color light-up RNA sensor for ratiometric imaging of miRNAs using Mango as a...

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Published inAnalytical chemistry (Washington) Vol. 93; no. 4; pp. 2534 - 2540
Main Authors Dou, Cai-Xia, Liu, Chaoyang, Ying, Zhan-Ming, Dong, Wanrong, Wang, Fenglin, Jiang, Jian-Hui
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 02.02.2021
Subjects
Biosensing Techniques
Cell fate
Cell Line
Cell lines
Cells (biology)
Chemistry
Color
Fluorescence
Gene expression
Genetic code
Genetic Engineering - methods
Humans
Imaging
Luminescent Proteins - genetics
Mangoes
MicroRNAs
MicroRNAs - chemistry
miRNA
Modules
Molecular Imaging - methods
Optical Imaging - methods
Ribonucleic acid
RNA
RNA - chemistry
Sensors
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Summary:MicroRNAs (miRNAs) play essential roles in regulating gene expression and cell fate. However, it remains a great challenge to image miRNAs with high accuracy in living cells. Here, we report a novel genetically encoded dual-color light-up RNA sensor for ratiometric imaging of miRNAs using Mango as an internal reference and SRB2 as the sensor module. This genetically encoded sensor is designed by expressing a splittable fusion of the internal reference and sensor module under a single promoter. This design strategy allows synchronous expression of the two modules with negligible interference. Live cell imaging studies reveal that the genetically encoded ratiometric RNA sensor responds specifically to mir-224. Moreover, the sensor-to-Mango fluorescence ratios are linearly correlated with the concentrations of mir-224, confirming their capability of determining mir-224 concentrations in living cells. Our genetically encoded light-up RNA sensor also enables ratiometric imaging of mir-224 in different cell lines. This strategy could provide a versatile approach for ratiometric imaging of intracellular RNAs, affording powerful tools for interrogating RNA functions and abundance in living cells.
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ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.0c04588