Journal article
Indexing TNF-α gene expression using a gene-targeted reporter cell line
BMC biology, Vol.7(1), pp.8-8
02/16/2009
DOI: 10.1186/1741-7007-7-8
PMCID: PMC2657777
PMID: 19220876
Abstract
Background: Current cell-based drug screening technologies utilize randomly integrated reporter genes to index transcriptional activity of an endogenous gene of interest. In this context, reporter expression is controlled by known genetic elements that may only partially capture gene regulation and by unknown features of chromatin specific to the integration site. As an alternative technology, we applied highly efficient gene-targeting with recombinant adeno-associated virus to precisely integrate a luciferase reporter gene into exon 1 of the HeLa cell tumor necrosis factor-alpha (TNF-alpha) gene. Drugs known to induce TNF-alpha expression were then used to compare the authenticity of gene-targeted and randomly integrated transcriptional reporters.
Results: TNF-alpha-targeted reporter activity reflected endogenous TNF-alpha mRNA expression, whereas randomly integrated TNF-alpha reporter lines gave variable expression in response to transcriptional and epigenetic regulators. 5,6-Dimethylxanthenone-4-acetic acid (DMXAA), currently used in cancer clinical trials to induce TNF-alpha gene transcription, was only effective at inducing reporter expression from TNF-alpha gene-targeted cells.
Conclusion: We conclude that gene-targeted reporter cell lines provide predictive indexing of gene transcription for drug discovery.
Details
- Title: Subtitle
- Indexing TNF-α gene expression using a gene-targeted reporter cell line
- Creators
- Ziying Yan - Department of Anatomy and Cell Biology, Carver College of Medicine, University of Iowa, Iowa City, IA, USADiana Lei-Butters - Department of Anatomy and Cell Biology, Carver College of Medicine, University of Iowa, Iowa City, IA, USAJohn F Engelhardt - Department of Anatomy and Cell Biology, Carver College of Medicine, University of Iowa, Iowa City, IA, USAGregory H Leno - Department of Anatomy and Cell Biology, Carver College of Medicine, University of Iowa, Iowa City, IA, USA
- Resource Type
- Journal article
- Publication Details
- BMC biology, Vol.7(1), pp.8-8
- DOI
- 10.1186/1741-7007-7-8
- PMID
- 19220876
- PMCID
- PMC2657777
- NLM abbreviation
- BMC Biol
- ISSN
- 1741-7007
- eISSN
- 1741-7007
- Publisher
- BioMed Central
- Language
- English
- Date published
- 02/16/2009
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
- Record Identifier
- 9984025303902771
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