Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 60, Issue 45, Pages 24070-24074Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202111170
Keywords
(p)ppGpp; biosensors; fluorogenic RNA; guanosine tetraphosphate; live-cell imaging
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Funding
- UMass Amherst start-up grant
- NIH [R01AI136789, R35GM130320]
- NSF CAREER
- Sloan Research Fellowship
- Camille Dreyfus Teacher-Scholar Award
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A RNA-based fluorescent sensor has been developed for live-cell imaging of (p)ppGpp, allowing direct monitoring of cellular accumulation and dynamics of (p)ppGpp levels. These sensors provide unprecedented information on cell-to-cell variation and can be broadly adapted to study bacterial stringent response.
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.
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