19. Bright Red Bioluminescence from Semisynthetic NanoLuc (sNLuc)
    Russo, F.; Civili, B.; Winssinger, N.
    ACS Chem. Biol. 2024, 19, 1035-1039

Red-shifted bioluminescence is highly desirable for diagnostic and imaging applications. Herein, we report a semisynthetic NanoLuc (sNLuc) based on complementation of a split NLuc (LgBiT) with a synthetic peptide (SmBiT) functionalized with a fluorophore for BRET emission. We observed exceptional BRET ratios with diverse fluorophores, notably in the red (I₆₇₄/I₄₅₀ > 14), with a brightness that is sufficient for naked eye detection in blood or through tissues. To exemplify its utility, LgBiT was fused to a miniprotein that binds HER2 (affibody, Z_HER2), and the selective detection of HER2⁺ SK-BR-3 cells over HER2^– HeLa cells was demonstrated.

DOI : 10.1021/acschembio.4c00033