Rapid discovery and evolution of nanosensors containing fluorogenic amino acids.

Autor:	Kuru E; Department of Genetics, Harvard Medical School, Boston, MA, USA. erkin_kuru@hms.harvard.edu.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA. erkin_kuru@hms.harvard.edu., Rittichier J; Department of Genetics, Harvard Medical School, Boston, MA, USA.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; EnPlusOne Biosciences Inc., Watertown, MA, USA., de Puig H; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Institute for Medical Engineering and Science and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA., Flores A; Department of Genetics, Harvard Medical School, Boston, MA, USA.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA., Rout S; Department of Genetics, Harvard Medical School, Boston, MA, USA.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA., Han I; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA., Reese AE; IRR Chemistry Hub and Centre for Inflammation Research, Institute for Regeneration and Repair, The University of Edinburgh, Edinburgh, UK., Bartlett TM; Department of Microbiology, Harvard Medical School, Boston, MA, USA.; Division of Genetics, Wadsworth Center, New York State Department of Health, Albany, NY, USA., De Moliner F; IRR Chemistry Hub and Centre for Inflammation Research, Institute for Regeneration and Repair, The University of Edinburgh, Edinburgh, UK., Bernier SG; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA., Galpin JD; Department of Molecular Physiology and Biophysics, The University of Iowa, Iowa City, IA, USA., Marchand J; Department of Genetics, Harvard Medical School, Boston, MA, USA.; Department of Chemical Engineering, University of Washington, Seattle, WA, USA., Bedell W; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA., Robinson-McCarthy L; Department of Genetics, Harvard Medical School, Boston, MA, USA., Ahern CA; Department of Molecular Physiology and Biophysics, The University of Iowa, Iowa City, IA, USA., Bernhardt TG; Department of Microbiology, Harvard Medical School, Boston, MA, USA.; Howard Hughes Medical Institute, Boston, MA, USA., Rudner DZ; Department of Microbiology, Harvard Medical School, Boston, MA, USA., Collins JJ; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.; Institute for Medical Engineering and Science and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.; Infectious Disease and Microbiome Program, Broad Institute of MIT and Harvard, Cambridge, MA, 02142, USA., Vendrell M; IRR Chemistry Hub and Centre for Inflammation Research, Institute for Regeneration and Repair, The University of Edinburgh, Edinburgh, UK. marc.vendrell@ed.ac.uk., Church GM; Department of Genetics, Harvard Medical School, Boston, MA, USA. gchurch@genetics.med.harvard.edu.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA. gchurch@genetics.med.harvard.edu.
Jazyk:	angličtina
Zdroj:	Nature communications [Nat Commun] 2024 Sep 05; Vol. 15 (1), pp. 7531. Date of Electronic Publication: 2024 Sep 05.
DOI:	10.1038/s41467-024-50956-z
Abstrakt:	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. (© 2024. The Author(s).)
Databáze:	MEDLINE
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