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Use of cDNA microarrays to probe and understand the toxicological consequences of altered gene expression
1AstraZeneca Central Toxicology Laboratory, Alderley Park, Cheshire, UK. bill.pennie@ctl.zeneca.com
Abstract:
Genomic sciences offer the ability to measure quantitative modulation of transcription in cells and tissues under a wide variety of conditions. We have developed a series of custom cDNA microarrays specifically to investigate toxicity processes. Around 600 marker genes for toxicity were selected and representative cDNA clones were obtained, amplified and purified by polymerase chain reaction (PCR), before being immobilised on nylon membranes. A detailed database on biochemical function, role in disease and allelic variation has been assembled for each gene. Applications in our laboratory include mechanistic investigation of a number of toxic endpoints such as hepatotoxicity and endocrine disruption.
Insights
Researchers created custom cDNA microarrays to study toxicity. These tools analyze gene expression changes, aiding in understanding toxic effects like liver damage and endocrine disruption.
Area of Science:
- Genomic sciences
- Toxicology
- Molecular biology
Background:
- Genomic sciences enable quantitative measurement of gene transcription modulation.
- Understanding cellular responses to various conditions is crucial.
- Toxicity processes require detailed investigation at the molecular level.
Purpose of the Study:
- To develop custom cDNA microarrays for investigating toxicity.
- To create a comprehensive database for selected toxicity marker genes.
- To apply these tools for mechanistic investigations of toxic endpoints.
Main Methods:
- Selection and validation of approximately 600 marker genes for toxicity.
- Amplification and purification of complementary DNA (cDNA) clones using polymerase chain reaction (PCR).
- Immobilization of purified cDNA clones onto nylon membranes to create microarrays.
- Assembly of a detailed database including biochemical function, disease relevance, and allelic variation for each gene.
Main Results:
- Development of custom cDNA microarrays tailored for toxicity studies.
- Creation of a curated database for 600 toxicity-associated genes.
- Successful application of microarrays in mechanistic investigations.
Conclusions:
- Custom cDNA microarrays are effective tools for studying toxicity.
- The developed gene database enhances the understanding of toxicity mechanisms.
- These methods facilitate the investigation of toxic endpoints like hepatotoxicity and endocrine disruption.