Live‐Cell Imaging of Guanosine Tetra‐ and Pentaphosphate (p)ppGpp with RNA‐based Fluorescent Sensors

Guanosine tetra‐ and pentaphosphate, (p)ppGpp, are important alarmone nucleotides that regulate bacterial survival in stressful environment. A direct detection of (p)ppGpp in living cells is critical for our understanding of the mechanism of bacterial stringent response. However, it is still challen...

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Bibliographic Details
Published inAngewandte Chemie International Edition Vol. 60; no. 45; pp. 24070 - 24074
Main Authors Sun, Zhining, Wu, Rigumula, Zhao, Bin, Zeinert, Rilee, Chien, Peter, You, Mingxu
Format Journal Article
LanguageEnglish
Published Germany Wiley Subscription Services, Inc 02.11.2021
EditionInternational ed. in English
Subjects
(p)ppGpp
Aptamers
Bacteria
biosensors
Broccoli
Escherichia coli - cytology
Fluorescence
Fluorescent Dyes
fluorogenic RNA
Genetic code
Guanosine
Guanosine Pentaphosphate
guanosine tetraphosphate
Guanosines
live-cell imaging
Nucleotides
Optical Imaging
Ribonucleic acid
RNA
Sensors
Spectrometry, Fluorescence
Stringent response
fluorogenic RNA
biosensors
(p)ppGpp
guanosine tetraphosphate
live-cell imaging
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Summary:Guanosine tetra‐ and pentaphosphate, (p)ppGpp, are important alarmone nucleotides that regulate bacterial survival in stressful environment. A direct detection of (p)ppGpp in living cells is critical for our understanding of the mechanism of bacterial stringent response. However, it is still challenging to image cellular (p)ppGpp. Here, we report RNA‐based fluorescent sensors for the live‐cell imaging of (p)ppGpp. Our sensors are engineered by conjugating a recently identified (p)ppGpp‐specific riboswitch with a fluorogenic RNA aptamer, Broccoli. These sensors can be genetically encoded and enable direct monitoring of cellular (p)ppGpp accumulation. Unprecedented information on cell‐to‐cell variation and cellular dynamics of (p)ppGpp levels is now obtained under different nutritional conditions. These RNA‐based sensors can be broadly adapted to study bacterial stringent response. Genetically encodable fluorescent RNA sensors are engineered to detect (p)ppGpp in live cells. Under nutrient starvation, cells produce and accumulate (p)ppGpp as a response, which can now be observed using these sensors.
Bibliography:https://www.biorxiv.org/content/10.1101/2021.05.12.443921v1
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A previous version of this manuscript has been deposited on a preprint server
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.202111170