期刊
NATURE COMMUNICATIONS
卷 11, 期 1, 页码 -出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/s41467-020-17228-y
关键词
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资金
- Cell Analysis Facility Flow Cytometry and Imaging Core in the Department of Immunology at the University of Washington
- Public Health Service, National Research Service Award from the National Institute of General Medical Sciences [T32GM007270]
- Seattle ARCS foundation
- University of Washington/Fred Hutchinson Cancer Research Center Viral Pathogenesis Training Program [2T32AI083203]
- University of Washington Medical Scientist Training Program [2T32GM007266]
- Ruth L. Kirschstein Predoctoral Fellowship [1F30CA239659-01A1]
- National Institutes of Health [1R21AI137758-01, 1R21AI153820-01]
Cyclic dinucleotides (CDNs) are second messengers conserved across all three domains of life. Within eukaryotes they mediate protective roles in innate immunity against malignant, viral, and bacterial disease, and exert pathological effects in autoimmune disorders. Despite their ubiquitous role in diverse biological contexts, CDN detection methods are limited. Here, using structure guided design of the murine STING CDN binding domain, we engineer a Forster resonance energy transfer (FRET) based biosensor deemed BioSTING. Recombinant BioSTING affords real-time detection of CDN synthase activity and inhibition. Expression of BioSTING in live human cells allows quantification of localized bacterial and eukaryotic CDN levels in single cells with low nanomolar sensitivity. These findings establish BioSTING as a powerful kinetic in vitro platform amenable to high throughput screens and as a broadly applicable cellular tool to interrogate the temporal and spatial dynamics of CDN signaling in a variety of infectious, malignant, and autoimmune contexts. Cyclic dinucleotides are conserved second messengers but current detection methods are limited. Here the authors engineer a Forster resonance energy transfer (FRET) based biosensor, BioSTING, which gives real-time in vitro detection of these nucleotides.
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