Hypoxia-Induced Reporter Genes with Different Half-Lives.

Abstract

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.

• Authors: Balaji Krishnamachary, Pierre Danhier, Samata Kakkad, Santosh K Bharti, Zaver M Bhujwalla
• Journal: Methods in molecular biology (Clifton, N.J.) (Methods Mol Biol) Vol. 1790 Pg. 113-125 ( 2018) ISSN: 1940-6029 [Electronic] United States
• PMID: 29858787 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• enhanced green fluorescent protein
• Green Fluorescent Proteins
• Luciferases

Topics

• Animals
• Gene Expression Regulation, Neoplastic
• Genes, Reporter
• Green Fluorescent Proteins (genetics, metabolism)
• Half-Life
• Heterografts
• Humans
• Hypoxia (physiopathology)
• Luciferases (genetics, metabolism)
• Male
• Mice
• Mice, SCID
• Molecular Imaging (methods)
• Neoplasm Transplantation
• Prostatic Neoplasms (genetics, metabolism, pathology)
• Tumor Cells, Cultured