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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Biosynthetic Lanthanide-Luminescent Mini-Proteins Using Genetic Code Expansion.

Edan Habel1, Haocheng Qianzhu1, Elwy H Abdelkader2

  • 1Research School of Chemistry, Australian National University , Canberra, ACT 2601, Australia.

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|March 31, 2026
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Summary
This summary is machine-generated.

Noncanonical amino acids enhance lanthanide luminescence in proteins. Computational design created a new mini-protein (RF2) with nanomolar binding affinity and improved terbium luminescence, demonstrating potential for bright, genetically encoded luminescent systems.

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Area of Science:

  • Biochemistry
  • Biophysics
  • Protein Engineering

Background:

  • Noncanonical amino acids (ncAA) show promise as light-harvesting antennae for lanthanide luminescence.
  • Understanding antenna-lanthanide interactions is key to designing brighter luminescent proteins.

Purpose of the Study:

  • To investigate antenna-lanthanide interactions for designing bright luminescent proteins.
  • To develop a computationally designed mini-protein with high lanthanide-binding affinity and luminescence.

Main Methods:

  • Computational protein design to create the RF2 scaffold.
  • Incorporation of 6-azatryptophan (6AW) as a noncanonical amino acid antenna.
  • Characterization of lanthanide luminescence enhancement (Tb3+, Eu3+, Dy3+, Sm3+) in the designed protein.

Main Results:

  • A novel protein, RF2, was designed with nanomolar lanthanide-binding affinity.
  • RF2 exhibited over a 2-fold increase in terbium(III) luminescence compared to lanthanide-binding tags (LBT).
  • The RF2 scaffold with 6-azatryptophan (6AW) achieved a 10-fold enhancement of europium(III) luminescence *in vivo* and sensitized other lanthanides.

Conclusions:

  • *De novo* protein design can yield highly luminescent lanthanide-binding mini-proteins.
  • Genetically encoded noncanonical amino acids serve as effective antennas for lanthanide luminescence.
  • The RF2 6AW system offers a powerful platform for developing novel luminescent biological tools.