Noninvasive imaging of cationic lipid-mediated delivery of optical and PET reporter genes in living mice.
Gene therapy involves the safe and effective delivery of one or more genes of interest to target cells in vivo. The advantages of using nonviral delivery systems include ease of preparation, low toxicity, and weak immunogenicity. Nonviral delivery methods, when combined with a noninvasive, clinically applicable imaging assay, will greatly aid in the optimization of gene therapy approaches for cancer. We demonstrate cationic lipid-mediated noninvasive monitoring of reporter gene expression of firefly (Photinus pyralis) luciferase (fl) and a mutant herpes simplex virus type I thymidine kinase (HSV1-sr39tk, tk) in living mice using a cooled charge coupled device (CCD) camera and positron emission tomography (PET), respectively. We observe a high level of fl and tk reporter gene expression predominantly in the lungs after a single injection of the extruded DOTAP:cholesterol DNA liposome complexes by way of the tail vein, seen to be time- and dose-dependent. We observe a good correlation between the in vivo bioluminescent signal and the ex vivo firefly luciferase enzyme (FL) activity in different organs. We further demonstrate the feasibility of noninvasively imaging both optical and PET reporter gene expression in the same animal using the CCD camera and microPET, respectively.
Iyer
Meera
M
The Crump for Molecular Imaging, Baylor College of Medicine, Houston, Texas 77030, USA.
Berenji
Manijeh
M
Templeton
Nancy S
NS
Gambhir
Sanjiv S
SS
D000818:Animals; D004247:DNA / Q000378:metabolism*; D018628:Gene Dosage; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000502:physiology*; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism*; D008168:Lung / Q000378:metabolism; D051379:Mice; D014055:Tomography, Emission-Computed
D008081:Liposomes; D004247:DNA