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Brighter NIR Bioluminescence System for Mammalian Cell Bioimaging Based on Engineered Railroadworm Luciferase and 6′-Aminoluciferin Analogues

Year: 2026

Type: article

Abstract: High Resolution Image Download MS PowerPoint Slide Luciferases have been extensively used as reporter genes for bioimaging purposes. For bioimaging in live animals, however, far-red- and NIR-emitting systems are better suited. Despite many efforts to develop such NIR-emitting systems, currently only a few systems have been commercialized, including AkaLuc/akalumine, which emits at ∼650 nm. Departing from the natural red-emitting luciferase of Phrixotrix hirtus railroadworm in combination with 6′-(1-pyrrolidinyl)luciferin (N5), we previously reported the development of a first efficient far-red-emitting system (650 nm). Here, we further improved this system, developing a luciferase that emits far-red bioluminescence with the natural substrate d -luciferin (630 nm), and a more efficient NIR bioluminescence with 6′-amino analogues (650–664 nm). The combination of this luciferase with N5 displayed superior properties compared with the commercial akalumine/AkaLuc system (645 nm), with high affinity for N5, higher catalytic efficiency and thermostability, more sustained luminescence, and more red-shifted spectrum (660 nm). COS-1 fibroblasts expressing this luciferase/N5 combination also displayed a brighter BL signal and more red-shifted spectrum when compared with the commercial akalumine/AkaLuc system, providing a promising NIR system for mammalian BL imaging.

Source: Chemical & Biomedical Imaging

DOI: 10.1021/cbmi.5c00163

Authors: Gabriel F. Pelentir, Vanessa R. Bevilaqua, Michio Kakiuchi, Takashi Hirano, Vadim R. Viviani

Institutions: Universidade Federal de São Carlos, Pontifícia Universidade Católica de São Paulo, University of Electro-Communications

Language: English

Topic: bioluminescence and chemiluminescence research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: diamond

Funders: Fundação de Amparo à Pesquisa do Estado de São Paulo, Conselho Nacional de Desenvolvimento Científico e Tecnológico, Japan Society for the Promotion of Science