Journal article
Rapid discovery and evolution of nanosensors containing fluorogenic amino acids
Nature communications, Vol.15(1), 7531
09/05/2024
DOI: 10.1038/s41467-024-50956-z
PMCID: PMC11377706
PMID: 39237489
Abstract
Binding-activated optical sensors are powerful tools for imaging, diagnostics, and biomolecular sensing. However, biosensor discovery is slow and requires tedious steps in rational design, screening, and characterization. Here we report on a platform that streamlines biosensor discovery and unlocks directed nanosensor evolution through genetically encodable fluorogenic amino acids (FgAAs). Building on the classical knowledge-based semisynthetic approach, we engineer ~15 kDa nanosensors that recognize specific proteins, peptides, and small molecules with up to 100-fold fluorescence increases and subsecond kinetics, allowing real-time and wash-free target sensing and live-cell bioimaging. An optimized genetic code expansion chemistry with FgAAs further enables rapid (~3 h) ribosomal nanosensor discovery via the cell-free translation of hundreds of candidates in parallel and directed nanosensor evolution with improved variant-specific sensitivities (up to ~250-fold) for SARS-CoV-2 antigens. Altogether, this platform could accelerate the discovery of fluorogenic nanosensors and pave the way to modify proteins with other non-standard functionalities for diverse applications.Binding-activated optical sensors are powerful tools, but their development can be slow and laborious. Here, authors introduce a platform to expedite biosensor discovery and evolution using genetically encodable fluorogenic amino acids.
Details
- Title: Subtitle
- Rapid discovery and evolution of nanosensors containing fluorogenic amino acids
- Creators
- Erkin Kuru - Harvard UniversityJonathan Rittichier - Harvard UniversityHelena de Puig - Massachusetts Institute of TechnologyAllison Flores - Harvard UniversitySubhrajit Rout - Harvard UniversityIsaac Han - Harvard UniversityAbigail E Reese - Centre for Inflammation ResearchThomas M Bartlett - Harvard UniversityFabio De Moliner - Centre for Inflammation ResearchSylvie G Bernier - Harvard UniversityJason D Galpin - University of IowaJorge Marchand - Harvard UniversityWilliam Bedell - Harvard UniversityLindsey Robinson-McCarthy - Harvard Medical SchoolChristopher A Ahern - University of IowaThomas G Bernhardt - Harvard UniversityDavid Z Rudner - Harvard UniversityJames J Collins - Massachusetts Institute of TechnologyMarc Vendrell - Centre for Inflammation ResearchGeorge M Church - Harvard University
- Resource Type
- Journal article
- Publication Details
- Nature communications, Vol.15(1), 7531
- Publisher
- Nature Publishing Group
- DOI
- 10.1038/s41467-024-50956-z
- PMID
- 39237489
- PMCID
- PMC11377706
- ISSN
- 2041-1723
- eISSN
- 2041-1723
- Grant note
- Wyss Institute Validation Project: DE-FG02-02ER63445 US Department of Energy: 771443
This work was supported by the Life Sciences Research Foundation Fellowship awarded to E.K. and Star Friedman Challenge and the Wyss Institute Validation Project awarded to G.M.C. and E.K. Work in the Church laboratory was supported by the US Department of Energy Grant DE-FG02-02ER63445. M.V. acknowledges funds from an ERC Consolidator Grant (DYNAFLUORS, 771443). We thank A. Kruse, M. Schubert, A. Chiappino-Pepe, and T.M. Wannier, for helpful discussions about this project; M. Lipstein and G. Kuziel for their advice on mRNA display; and G. Chao for his help with sensor dynamics experiments. We also thank the technical support from the Center for Macromolecular Interactions at Harvard Medical School, as well as the Harvard Medical School Core facilities for Taplin Mass Spectrometry and the Imaging/Microscopy (MICRON) Center.
- Language
- English
- Date published
- 09/05/2024
- Academic Unit
- Molecular Physiology and Biophysics; Iowa Neuroscience Institute
- Record Identifier
- 9984702559402771
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