An improved pathway for autonomous bioluminescence imaging in eukaryotes

Abstract

The discovery of the bioluminescence pathway in the fungus Neonothopanus nambi enabled engineering of eukaryotes with self-sustained luminescence. However, the brightness of luminescence in heterologous hosts was limited by performance of the native fungal enzymes. Here we report optimized versions of the pathway that enhance bioluminescence by one to two orders of magnitude in plant, fungal and mammalian hosts, and enable longitudinal video-rate imaging.

Improvement in Tol2 transposon for efficient large-cargo capacity transgene applications in cultured cells and zebrafish (<i>Danio rerio</i>)

“…These findings position Tol2 transposons as viable alternatives for delivering large DNA cargos in plasmids, especially where multiple genetic elements need to be incorporated, such as the whole luciferin synthetic pathway for bioluminescence. Despite challenges in detecting nnLuz luminescence in transgenic zebrafish, further improvements in enzyme activity (Beregovaya et al, 2021) or synthetic pathways (Shakhova et al, 2024) may unlock its full potential for cellular processes in vivo. Furthermore, Tol2 could facilitate the integration of multivalent chimeric antigen receptors into T cells, offering new avenues in immunotherapy for targeting solid tumors with neoantigens (Riley et al, 2019).…”