TY - JOUR
AU - Kuru, Erkin
AU - Rittichier, Jonathan
AU - Flores, Allison
AU - Rout, Subhrajit
AU - Han, Isaac
AU - Reese, Abigail E.
AU - Bartlett, Thomas M.
AU - Bernier, Sylvie G.
AU - Galpin, Jason D.
AU - Marchand, Jorge
AU - Bedell, William
AU - Robinson-McCarthy, Lindsey
AU - Ahern, Christopher A.
AU - Bernhardt, Thomas G.
AU - Rudner, David Z.
AU - Collins, James J.
AU - Vendrell, Marc
AU - Church, George M.
AU - de Puig, Helena
AU - de Moliner, Fabio
PY - 2024
DA - September
TI - Rapid discovery and evolution of nanosensors containing fluorogenic amino acids
JO - Nature Communications
VL - 15
DO - https://doi.org/10.1038/s41467-024-50956-z
AB - 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.
PB - Springer Nature
UR - https://www.research.ed.ac.uk/en/publications/69c96275-f15c-4bf2-82bb-a756399e3e30
KW - Coronavirus (COVID-19)
ER