Hypoxia-Induced Reporter Genes with Different Half-Lives
Open accessAbstract
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.
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The relatively short half-life of luciferase may have an impact on the results, preventing the detection of differences in cumulative transgene expression.
“The relatively short half-life of luciferase may have an impact on the results [43], preventing the detection of differences in the cumulative transgene expression.”
From Untranslated Region Sequences and the Efficacy of mRNA Vaccines against Tuberculosis (2024) · cites this paper as [43]
The Introduction states that luciferase has a half-life of only 2 hours, and the paper contrasts short-half-life luciferase with long-half-life GFP reporters. It does not state that luciferase's half-life prevented detection of differences in cumulative transgene expression; that proposed effect on the citing study's results is not supported by this source. (LLM, conf=0.82)
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Cited by
Sorted by citations
Untranslated Region Sequences and the Efficacy of mRNA Vaccines against Tuberculosis
Reshetnikov, V., Terenin, I., Shepelkova, G., Yeremeev, V., Kolmykov, S., Nagornykh, M., Kolosova, E., Sokolova, T., Zaborova, O., Kukushkin, I., et al. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms25020888 · Cited by 33 · see citing works
- Authors
- Krishnamachary, B., Danhier, P., Kakkad, S., Bharti, S. K., & Bhujwalla, Z. M.
- Affiliations
- Johns Hopkins University (US)
- Corresponding author
- Krishnamachary, Balaji
- Venue
- Methods in Molecular Biology
- Publisher
- Springer New York
- Published
- 2018
- Volume
- 1790
- Pages
- 113-125
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- PMC
- PMC7990048
- OpenAlex
- W2806899619
- Cited by
- 4
- References
- 0
- Citations by year
- 2025: 2 · 2024: 1 · 2018: 1
- FWCI
- 0.32
- Type
- book chapter
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health
- National Cancer Institute (R01CA082337, P50CA103175, R01CA073850, R35CA209960, P30CA006973)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › bioluminescence and chemiluminescence research
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Optical Imaging and Spectroscopy Techniques
- Keywords
- enhanced green fluorescent protein, luciferase, prostate cancer xenografts, PC-3 cells, tumor hypoxia, in vivo imaging
- MeSH terms
- Animals, Hypoxia, Half-Life, Humans, Luciferases, Male, Neoplasm Transplantation, Prostatic Neoplasms, Tumor Cells, Cultured, Gene Expression Regulation, Neoplastic, Mice, SCID, Genes, Reporter, Green Fluorescent Proteins, Mice, Molecular Imaging, Heterografts
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