Optical imaging of Renilla luciferase, synthetic Renilla luciferase, and firefly luciferase reporter gene expression in living mice

J Biomed Opt. 2004 May-Jun;9(3):578-86. doi: 10.1117/1.1647546.

Abstract

We have recently demonstrated that Renilla luciferase (Rluc) is a promising bioluminescence reporter gene that can be used for noninvasive optical imaging of reporter gene expression in living mice, with the aid of a cooled charged couple device (CCD) camera. In the current study, we explore the expression of a novel synthetic Renilla luciferase reporter gene (hRluc) in living mice, which has previously been reported to be a more sensitive reporter than native Rluc in mammalian cells. We explore the strategies of simultaneous imaging of both Renilla luciferase enzyme (RL) and synthetic Renilla luciferase enzyme (hRL):coelenterazine (substrate for RL/hRL) in the same living mouse. We also demonstrate that hRL:coelenterazine can yield a higher signal when compared to Firefly luciferase enzyme (FL): D-Luciferin, both in cell culture studies and when imaged from cells at the surface and from lungs of living mice. These studies demonstrate that hRluc should be a useful primary reporter gene with high sensitivity when used alone or in conjunction with other bioluminescence reporter genes for imaging in living rodents.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.
  • Validation Study

MeSH terms

  • Animals
  • Anthozoa / enzymology*
  • COS Cells
  • Cell Line
  • Chlorocebus aethiops
  • Coleoptera / enzymology*
  • Firefly Luciferin / pharmacokinetics*
  • Genes, Reporter / genetics
  • Glioma / metabolism
  • Humans
  • Imidazoles*
  • Isoenzymes
  • Kidney / metabolism
  • Luciferases / genetics
  • Luciferases / metabolism*
  • Metabolic Clearance Rate
  • Mice
  • Microscopy, Fluorescence / methods*
  • Neuroblastoma / metabolism
  • Organ Specificity
  • Pyrazines / pharmacokinetics*
  • Rats
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Substrate Specificity
  • Tissue Distribution

Substances

  • Imidazoles
  • Isoenzymes
  • Pyrazines
  • coelenterazine
  • Firefly Luciferin
  • Luciferases