Red-emitting chimeric firefly luciferase for in vivo imaging in low ATP cellular environments.

ABSTRACT: Beetle luciferases have been adapted for live cell imaging where bioluminescence is dependent on the cellular availability of ATP, O₂, and added luciferin. Previous Photinus pyralis red-emitting variants with high K_m values for ATP have performed disappointingly in live cells despite having much higher relative specific activities than enzymes like Click Beetle Red (CBR). We engineered a luciferase variant PLR3 having a K_m value for ATP similar to CBR and ∼2.6-fold higher specific activity. The red-emitting PLR3 was ∼2.5-fold brighter than CBR in living HEK293T and HeLa cells, an improvement consistent with the importance of the K_m value in low ATP environments.

SUBMITTER: Branchini BR 

PROVIDER: S-EPMC5730985 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature