Abstract
The luminescent reporter gene assay (LRGA) is arguably the most prominent type of reporter gene assay used in biomolecular and pharmaceutical development laboratories. Part of this popularity is due to the high signal associated with luciferases, the foundation of this technology. This feature makes them ideally suited for high throughput screening applications where potentially millions of chemical compounds can be analyzed in a given assay. Recent technical advancements that enhance signal stability of the luciferases along with development and commercialization of multiple forms of luciferases, their respective substrates, and improvements in expression vectors for reporter gene assay (RGA) applications have broadened their use. While the practical challenges related to the application of luminescent technology in a laboratory setting have been overcome, there remains much to do in laying a systematic approach towards the construction of RGAs, which are essential to the elucidation of the basic biology for genes of interest. This mini-review aims at giving a birds-eye view of the available luciferases, substrates and other luminescent technologies available and provides a general blueprint as well as practical considerations for constructing and interfacing RGAs with chemical biology and functional genomics for the elucidation of fundamental biological questions and for biomedical research.
Keywords: Luciferase Renilla, firefly, reporter gene assay (RGA), promoter, luciferin, high throughput screening, biomedical research, irradiation, Coelenterazine, co-enzyme, promoters, TATA box, cDNA
Combinatorial Chemistry & High Throughput Screening
Title: Seeing the Light: Luminescent Reporter Gene Assays
Volume: 14 Issue: 8
Author(s): Loren J. Miraglia, Frederick J. King and Robert Damoiseaux
Affiliation:
Keywords: Luciferase Renilla, firefly, reporter gene assay (RGA), promoter, luciferin, high throughput screening, biomedical research, irradiation, Coelenterazine, co-enzyme, promoters, TATA box, cDNA
Abstract: The luminescent reporter gene assay (LRGA) is arguably the most prominent type of reporter gene assay used in biomolecular and pharmaceutical development laboratories. Part of this popularity is due to the high signal associated with luciferases, the foundation of this technology. This feature makes them ideally suited for high throughput screening applications where potentially millions of chemical compounds can be analyzed in a given assay. Recent technical advancements that enhance signal stability of the luciferases along with development and commercialization of multiple forms of luciferases, their respective substrates, and improvements in expression vectors for reporter gene assay (RGA) applications have broadened their use. While the practical challenges related to the application of luminescent technology in a laboratory setting have been overcome, there remains much to do in laying a systematic approach towards the construction of RGAs, which are essential to the elucidation of the basic biology for genes of interest. This mini-review aims at giving a birds-eye view of the available luciferases, substrates and other luminescent technologies available and provides a general blueprint as well as practical considerations for constructing and interfacing RGAs with chemical biology and functional genomics for the elucidation of fundamental biological questions and for biomedical research.
Export Options
About this article
Cite this article as:
J. Miraglia Loren, J. King Frederick and Damoiseaux Robert, Seeing the Light: Luminescent Reporter Gene Assays, Combinatorial Chemistry & High Throughput Screening 2011; 14 (8) . https://dx.doi.org/10.2174/138620711796504389
DOI https://dx.doi.org/10.2174/138620711796504389 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
Call for Papers in Thematic Issues
Artificial Intelligence Methods for Biomedical, Biochemical and Bioinformatics Problems
Recently, a large number of technologies based on artificial intelligence have been developed and applied to solve a diverse range of problems in the areas of biomedical, biochemical and bioinformatics problems. By utilizing powerful computing resources and massive amounts of data, methods based on artificial intelligence can significantly improve the ...read more
Eco-friendly Agents for Biological Control of Pathogenic Diseases
The discovery of an alternative biological approach to disease management includes work on medicinal products derived from natural sources as a starting point for the development of eco-friendly agents for these diseases and the injuries they cause, as well as reducing human contact with hazardous chemicals and their residues. We ...read more
Emerging trends in diseases mechanisms, noble drug targets and therapeutic strategies: focus on immunological and inflammatory disorders
Recently infectious and inflammatory diseases have been a key concern worldwide due to tremendous morbidity and mortality world Wide. Recent, nCOVID-9 pandemic is a good example for the emerging infectious disease outbreak. The world is facing many emerging and re-emerging diseases out breaks at present however, there is huge lack ...read more
Exploring Spectral Graph Theory in Combinatorial Chemistry
Scope of the Thematic Issue: Combinatorial chemistry involves the synthesis and analysis of a large number of diverse compounds simultaneously. Traditional methods rely on brute force experimentation, which can be time-consuming and resource-intensive. Spectral Graph Theory, a branch of mathematics dealing with the properties of graphs in relation to the ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Transactivation of ErbB Receptors by Leptin in the Cardiovascular System: Mechanisms, Consequences and Target for Therapy
Current Pharmaceutical Design Strategies to Convert PACAP from a Hypophysiotropic Neurohormone Into a Neuroprotective Drug
Current Pharmaceutical Design ent-Kaurane Diterpenoids from Croton tonkinensis Induce Apoptosis in Colorectal Cancer Cells through the Phosphorylation of JNK Mediated by Reactive Oxygen Species and Dual-Specificity JNK Kinase MKK4
Anti-Cancer Agents in Medicinal Chemistry A Diabetes Treatment Strategy to Reduce the Risk of Cardiovascular Events: Clinical Benefits and Potential of Linagliptin
Immunology, Endocrine & Metabolic Agents in Medicinal Chemistry (Discontinued) Overcoming the Drug Resistance Problem with Second-Generation Tyrosine Kinase Inhibitors: From Enzymology to Structural Models
Current Medicinal Chemistry Current & Future Therapies of Erectile Dysfunction in Neurological Disorders
Recent Patents on CNS Drug Discovery (Discontinued) Effects of Iron Chelation in Osteosarcoma
Current Cancer Drug Targets Non-Small Cell Lung Cancer: From Cytotoxic Systemic Chemotherapy to Molecularly Targeted Therapy
Current Medicinal Chemistry - Anti-Cancer Agents Targeting AMPK Signaling Pathway to Overcome Drug Resistance for Cancer Therapy
Current Drug Targets Using Pharmacogenomic Tumor Profiling to Identify Biomarkers of 5-fluorouracil Response in Colorectal Cancer
Current Pharmacogenomics Novel Targets for Antiinflammatory and Antiarthritic Agents
Current Pharmaceutical Design Primary Cilia in Tumor Biology: The Primary Cilium as a Therapeutic Target in Cholangiocarcinoma
Current Drug Targets Oncomirs: From Tumor Biology to Molecularly Targeted Anticancer Strategies
Mini-Reviews in Medicinal Chemistry Implications of the Dominant Role of Transporters in Drug Uptake by Cells
Current Topics in Medicinal Chemistry A Medicinal Chemist’s Perspective Towards Structure Activity Relationship of Heterocycle Based Anticancer Agents
Current Topics in Medicinal Chemistry 4D-QSAR Models Applied to the Study of TGF- β1 Receptor Inhibitors
Current Topics in Medicinal Chemistry Anti-Cancer Potential of Some Commonly Used Drugs
Current Pharmaceutical Design Ferrous Ion Chelating Modification and Treatment of Iron-Deficiency Anemia of Exopolysaccharide from Lachnum sp.
Current Chemical Biology Surgery for Severe Obesity: Indications, Techniques, Mechanisms of Weight Loss and Diabetes Resolution
Immunology, Endocrine & Metabolic Agents in Medicinal Chemistry (Discontinued) Oncogene-Directed Therapies as Modulators of Cancer Coagulopathy, Angiogenesis and Tumor-Vascular Interface
Current Signal Transduction Therapy