Hypoxia-Induced Reporter Genes with Different Half-Lives
The utility of reporter genes has gained significant momentum over the last three decades. Reporter genes are used to understand the transcriptional activity of a gene both in vitro and in vivo, and in pathway analysis and drug screening for diseases involving protozoan parasites, and in anti-cancer drug developments. Here, using a human prostate cancer xenograft model (PC3), we describe a method to construct and validate hypoxia reporter genes with different half-lives. Using molecular biology and optical imaging techniques, we have validated the expression of long half-life enhanced green fluorescence protein (EGFP) expression and short half-life luciferase gene expression to report on the spatial and temporal evolution of hypoxia in vivo.
Key wordsBioluminescence Lentivirus Luciferase assay Hypoxia Hypoxia response elements (HRE) Reporter gene
This work was supported by National Institutes of Health R01 CA73850, R01 CA82337, and P50 CA103175.
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