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Fluorescent Nanoparticles for the Measurement of Ion Concentration in Biological Systems
Published on: July 4, 2011
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Rapid discovery and evolution of nanosensors containing fluorogenic amino acids
Erkin Kuru1,2, Jonathan Rittichier3,4,5, Helena de Puig4,6
1Department of Genetics, Harvard Medical School, Boston, MA, USA. erkin_kuru@hms.harvard.edu.
Nature Communications
|September 5, 2024
Summary
This study introduces a new platform using genetically encodable fluorogenic amino acids (FgAAs) to rapidly discover and evolve optical nanosensors. This accelerates the development of sensitive biosensors for real-time diagnostics and imaging.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Optical biosensors are crucial for imaging and diagnostics but their discovery is slow.
- Current methods involve tedious rational design, screening, and characterization processes.
Purpose of the Study:
- To develop a streamlined platform for rapid biosensor discovery and evolution.
- To engineer novel nanosensors using genetically encodable fluorogenic amino acids (FgAAs).
Main Methods:
- Utilized knowledge-based semisynthetic approach combined with genetic code expansion.
- Employed cell-free translation for parallel screening of hundreds of nanosensor candidates.
- Implemented directed evolution for optimizing nanosensor sensitivity.
Main Results:
- Engineered ~15 kDa nanosensors with up to 100-fold fluorescence increases and subsecond kinetics.
- Achieved rapid ribosomal nanosensor discovery (~3 hours) using FgAAs.
- Demonstrated improved variant sensitivities (up to ~250-fold) for SARS-CoV-2 antigens.
Conclusions:
- The developed platform significantly accelerates fluorogenic nanosensor discovery and evolution.
- This approach enables real-time, wash-free sensing and live-cell bioimaging.
- The technology holds potential for modifying proteins with non-standard functionalities for diverse applications.
